BNIP3L Antibody

Immunocytochemistry/ Immunofluorescence: BNIP3L Antibody [NBP1-88558]

Immunocytochemistry/Immunofluorescence: BNIP3L Antibody [NBP1-88558] - Staining of human cell line U-2 OS shows localization to nuclear speckles & mitochondria. Antibody staining is shown in green.

Immunohistochemistry-Paraffin: BNIP3L Antibody [NBP1-88558]

Immunohistochemistry-Paraffin: BNIP3L Antibody [NBP1-88558] - Staining of human pancreas shows low expression as expected.

Immunohistochemistry-Paraffin: BNIP3L Antibody [NBP1-88558]

Immunohistochemistry-Paraffin: BNIP3L Antibody [NBP1-88558] - Staining of human placenta shows high expression.

Western Blot: BNIP3L Antibody [NBP1-88558] -

Corticosterone affects NIX-dependent mitophagy through decreasing PGC1 alpha in vivo.a–f Mice exposed to vehicle, corticosterone (10 mg/kg), corticosterone with phorbol 12-myristate 13-acetate (PMA pretreatment, 200 μg/kg), or PMA alone for 7 days. a Slide samples for IHC immunostained with LAMP1 (green), TOMM20 (red), & DAPI (blue). Scale bars, 100 μm (magnification, ×200). n = 5. b The expressions of NIX, PTEN-induced kinase 1 (PINK1), & BCL2 interacting protein 3 (BNIP3) detected with WBt where beta-actin used as a loading control. n = 5. c Slide samples for IHC immunostained with synpatophysin (green), PSD95 (red), & DAPI (blue). Scale bars, 100 μm (magnification, ×200). n = 5. d Synaptophysin & PSD95 detected by WBt. Loading control is beta-actin. n = 5. e The mice subjected to Y-maze test to evaluate spatial memory function. n = 6. f The mice subjected to forced swim test to evaluate depression-like behavior. n = 5. g Vehicle or RU 486 (5 mg/kg) injected mice presented with/without corticosterone (10 mg/kg) for 3 days. The expressions of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1 alpha) & NIX visualized via WB. Loading control is beta-actin. n = 5. h The schematic model for mechanisms of inhibition in NIX-dependent mitophagy by glucocorticoid was shown. All blots & IF images representative. n = 5 or 6 from each animal with two technical replicates each in results of IHC & WBt. Quantitative data presented as a mean ± S.E.M. The representative images acquired by SRRF imaging system. Two-sided two-way ANOVA was conducted except Fig. 8b, data of which analyzed by two-sided unpaired student’s t test. ** indicates p < 0.01 versus control & ## indicates p < 0.01 versus corticosterone, respectively. Data provided as a Source data file. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33473105), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: BNIP3L Antibody [NBP1-88558] -

Western Blot: BNIP3L Antibody [NBP1-88558] - Role of PGC1 alpha in NIX-dependent mitophagy.a–e Nontargeting (NT) or GR siRNA was transfected to hippocampal neurons & SH-SY5Y cells for 24 h prior to corticosterone & cortisol for 12 h, respectively. a, b Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1 alpha) expression was detected in western blot where beta-actin was used as a loading control in both cell types. n = 5. c Colocalization of PGC1 alpha (red) & DAPI (blue) in hippocampal neurons was visualized with SRRF imaging system. Scale bars, 20 μm (magnification, ×1000). n = 5. d Colocalization of PGC1 alpha (green) & DAPI (blue) in SH-SY5Y was visualized with SRRF imaging system. Scale bars, 20 μm (magnification, ×1000). n = 5. e PGC1 alpha protein expressions in subcellular fraction samples were detected by western blotting. Lamin A/C & alpha-tubulin were used as a nuclear & cytosolic loading control, respectively. n = 5. f, g SH-SY5Y cells were transfected with pcDNA3.1/c-eGFP or pcDNA3.1/PPARGC1A-c-eGFP vector for 24 h prior to cortisol treatment for 24 h. f NIX expression was detected in western blot where beta-actin was used as a loading control. n = 5. g TOMM20 levels were detected by western blot. Loading control for western blot is beta-actin. n = 5. All blots & immunofluorescence images are representative. n = 5 from independent experiments with two technical replicates each. Quantitative data are presented as a mean ± S.E.M. Two-sided two-way ANOVA was conducted. ** indicates p < 0.01 versus control. #, ## indicates p < 0.05, p < 0.01 versus corticosterone in hippocampal neurons & cortisol in SH-SY5Y, respectively. Data are provided as a Source data file. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33473105), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: BNIP3L Antibody [NBP1-88558] -

Western Blot: BNIP3L Antibody [NBP1-88558] - GR-dependent downregulation of NIX expression via PGC1 alpha.a, b Nontargeting (NT) or GR siRNA was transfected to hippocampal neurons & SH-SY5Y cells for 24 h prior to corticosterone & cortisol for 24 h, respectively. NIX expression was detected in western blot where beta-actin was used as a loading control. n = 5. c SH-SY5Y cells were incubated with cortisol for 6 h. The mRNA expression levels of genes associated with nuclear receptors & coregulators were assessed by RT2 Profiler PCR array. Heat maps with hierarchical clustering were acquired by using the GeneGlobe Data analysis Center on Qiagen website. n = 3. d A thousand base pair upstream of the first codon of the PPARGC1A was described & the putative GRE binding sequence was emphasized with yellow labeling. e SH-SY5Y cells were incubated with cortisol for 6 h. DNA was immunoprecipitated with IgG, RNA polymerase (RNAPol), & glucocorticoid receptor (GR) antibody. The immunoprecipitation & input samples were amplified with primers of GAPDH & PPARGC1A gene. n = 5. All blots are representative. n = 3 or 5 from independent experiments with two technical replicates each, respectively. Quantitative data are presented as a mean ± S.E.M. Two-sided two-way ANOVA was conducted in Fig. 6a, b. Two-sided unpaired student’s t test was conducted in Fig. 6e. ** indicates p < 0.01 versus control. ## indicates p < 0.01 versus corticosterone in hippocampal neurons & cortisol in SH-SY5Y, respectively. Data are provided as a Source data file. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33473105), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: BNIP3L Antibody [NBP1-88558] -

Western Blot: BNIP3L Antibody [NBP1-88558] - GR-dependent downregulation of NIX expression via PGC1 alpha.a, b Nontargeting (NT) or GR siRNA was transfected to hippocampal neurons & SH-SY5Y cells for 24 h prior to corticosterone & cortisol for 24 h, respectively. NIX expression was detected in western blot where beta-actin was used as a loading control. n = 5. c SH-SY5Y cells were incubated with cortisol for 6 h. The mRNA expression levels of genes associated with nuclear receptors & coregulators were assessed by RT2 Profiler PCR array. Heat maps with hierarchical clustering were acquired by using the GeneGlobe Data analysis Center on Qiagen website. n = 3. d A thousand base pair upstream of the first codon of the PPARGC1A was described & the putative GRE binding sequence was emphasized with yellow labeling. e SH-SY5Y cells were incubated with cortisol for 6 h. DNA was immunoprecipitated with IgG, RNA polymerase (RNAPol), & glucocorticoid receptor (GR) antibody. The immunoprecipitation & input samples were amplified with primers of GAPDH & PPARGC1A gene. n = 5. All blots are representative. n = 3 or 5 from independent experiments with two technical replicates each, respectively. Quantitative data are presented as a mean ± S.E.M. Two-sided two-way ANOVA was conducted in Fig. 6a, b. Two-sided unpaired student’s t test was conducted in Fig. 6e. ** indicates p < 0.01 versus control. ## indicates p < 0.01 versus corticosterone in hippocampal neurons & cortisol in SH-SY5Y, respectively. Data are provided as a Source data file. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33473105), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: BNIP3L Antibody [NBP1-88558] -

Western Blot: BNIP3L Antibody [NBP1-88558] - Effects of hypoxia on mitophagy regulator expressions & mitophagy in UCB-hMSCs. (A) UCB-hMSCs incubated w/ various times of hypoxia (0–48 h). Cells stained w/ Mitotracker™. n = 6 (magnification, ×1,200). Scale bars, 50 µm. (B) The expressions of COX4 & beta-actin detected by western blot. n = 4. (C) UCB-hMSCs exposed to 24 h of normoxia or hypoxia. Cells immuno-stained w/ COX4 & LC3B-specific antibodies (magnification, ×600). Scale bars, 37.5 µm. (D) The mRNA expressions of PINK1, BNIP3, NIX & FUNDC1 analyzed by quantitative real-time PCR (qPCR). n = 5. (E) The protein expressions of PINK1, BNIP3, NIX & beta-actin assessed by western blot. n = 4. (F, G) siRNAs of PINK1, BNIP3, NIX or non-targeting (NT) transfected to UCB-hMSCs prior to hypoxia treatment for 24 h. COX4 & beta-actin expressions assessed by western blot. n = 4 (F). Cells immunostained w/ COX4 & PI. n = 3 (magnification, ×400). All scale bars, 50 µm. COX4 fluorescence intensity analyzed by luminometer. n = 5 (G). (H) BNIP3 siRNA transfected to UCB-hMSCs prior to hypoxia treatment for 24 h. Cells stained w/ Mitotracker™. n = 6 (magnification, ×1,200). Scale bars, 50 µm. (I) BNIP3, beta-tubulin & COX4 w/ cytosol & mitochondrial fractionized samples detected by western blot. (J) Cells incubated w/ hypoxia or normoxia for 24 h. Cells immuno-stained w/ BNIP3 & LC3B-specific antibodies. (magnification, ×600). Scale bars, 37.5 µm. Western blot data normalized by beta-actin, & qPCR data normalized by ACTB mRNA expression level. Quantitative data are presented as a mean ± S.E.M. All blot & confocal images are representative. *p < 0.05 versus control, #p < 0.05 versus hypoxia. Image collected & cropped by CiteAb from the following publication (https://linkinghub.elsevier.com/retrieve/pii/S2213231717303804), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: BNIP3L Antibody [NBP1-88558] -

Immunocytochemistry/ Immunofluorescence: BNIP3L Antibody [NBP1-88558] - The role of PA in BNIP3 silenced UCB-hMSC survival in the mouse skin wound healing model. (A) Mouse skin wound surgery with UCB-hMSC transplantation was performed as described in Section 2. Representative gross wound images were acquired at post injection days 0, 4, 8, 12. Skin wound sizes at day 8 were compared with wound size at day 0. n = 5. (B) Tissue slide samples were stained with hematoxylin & eosin. Low & high magnified histological gross images are shown in the left & right panels, respectively. Scale bars, 260 µm (magnification, ×40) & 100 µm (magnification, ×100). n = 5. (C) Representative images of blood vessels in skin wounds on day 12 (top panel). Vessel density was analyzed by using ImageJ program (bottom panel). n = 5. (D-F) Histological tissue samples were immuno-stained with CD31, alpha-SMA, & HNA-specific antibodies & PI for counterstaining. alpha-SMA & HNA-positive cells were visualized by confocal microscopy. The number of CD31 & alpha-SMA-positive cells in high power field (HPF), & the percentage of HNA-positive cells in total cells were analyzed by using Metamorph software. Scale bars, 100 µm (magnification, ×100). n = 5. Data are presented as a mean ± S.E.M. $p < 0.05 versus vehicle group, *p < 0.05 versus UCB-hMSC group given NT siRNA, #p < 0.05 versus UCB-hMSC group given NT siRNA with hypoxia pretreatment, @p < 0.05 versus UCB-hMSC group given BNIP3 siRNA with hypoxia pretreatment. Image collected & cropped by CiteAb from the following publication (https://linkinghub.elsevier.com/retrieve/pii/S2213231717303804), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: BNIP3L Antibody [NBP1-88558] -

Immunocytochemistry/ Immunofluorescence: BNIP3L Antibody [NBP1-88558] - The role of PA in BNIP3 silenced UCB-hMSC survival in the mouse skin wound healing model. (A) Mouse skin wound surgery with UCB-hMSC transplantation was performed as described in Section 2. Representative gross wound images were acquired at post injection days 0, 4, 8, 12. Skin wound sizes at day 8 were compared with wound size at day 0. n = 5. (B) Tissue slide samples were stained with hematoxylin & eosin. Low & high magnified histological gross images are shown in the left & right panels, respectively. Scale bars, 260 µm (magnification, ×40) & 100 µm (magnification, ×100). n = 5. (C) Representative images of blood vessels in skin wounds on day 12 (top panel). Vessel density was analyzed by using ImageJ program (bottom panel). n = 5. (D-F) Histological tissue samples were immuno-stained with CD31, alpha-SMA, & HNA-specific antibodies & PI for counterstaining. alpha-SMA & HNA-positive cells were visualized by confocal microscopy. The number of CD31 & alpha-SMA-positive cells in high power field (HPF), & the percentage of HNA-positive cells in total cells were analyzed by using Metamorph software. Scale bars, 100 µm (magnification, ×100). n = 5. Data are presented as a mean ± S.E.M. $p < 0.05 versus vehicle group, *p < 0.05 versus UCB-hMSC group given NT siRNA, #p < 0.05 versus UCB-hMSC group given NT siRNA with hypoxia pretreatment, @p < 0.05 versus UCB-hMSC group given BNIP3 siRNA with hypoxia pretreatment. Image collected & cropped by CiteAb from the following publication (https://linkinghub.elsevier.com/retrieve/pii/S2213231717303804), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: BNIP3L Antibody [NBP1-88558] -

Immunocytochemistry/ Immunofluorescence: BNIP3L Antibody [NBP1-88558] - The role of PA in BNIP3 silenced UCB-hMSC survival in the mouse skin wound healing model. (A) Mouse skin wound surgery with UCB-hMSC transplantation was performed as described in Section 2. Representative gross wound images were acquired at post injection days 0, 4, 8, 12. Skin wound sizes at day 8 were compared with wound size at day 0. n = 5. (B) Tissue slide samples were stained with hematoxylin & eosin. Low & high magnified histological gross images are shown in the left & right panels, respectively. Scale bars, 260 µm (magnification, ×40) & 100 µm (magnification, ×100). n = 5. (C) Representative images of blood vessels in skin wounds on day 12 (top panel). Vessel density was analyzed by using ImageJ program (bottom panel). n = 5. (D-F) Histological tissue samples were immuno-stained with CD31, alpha-SMA, & HNA-specific antibodies & PI for counterstaining. alpha-SMA & HNA-positive cells were visualized by confocal microscopy. The number of CD31 & alpha-SMA-positive cells in high power field (HPF), & the percentage of HNA-positive cells in total cells were analyzed by using Metamorph software. Scale bars, 100 µm (magnification, ×100). n = 5. Data are presented as a mean ± S.E.M. $p < 0.05 versus vehicle group, *p < 0.05 versus UCB-hMSC group given NT siRNA, #p < 0.05 versus UCB-hMSC group given NT siRNA with hypoxia pretreatment, @p < 0.05 versus UCB-hMSC group given BNIP3 siRNA with hypoxia pretreatment. Image collected & cropped by CiteAb from the following publication (https://linkinghub.elsevier.com/retrieve/pii/S2213231717303804), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: BNIP3L Antibody [NBP1-88558] -

Immunocytochemistry/ Immunofluorescence: BNIP3L Antibody [NBP1-88558] - The role of PA in BNIP3 silenced UCB-hMSC survival in the mouse skin wound healing model. (A) Mouse skin wound surgery with UCB-hMSC transplantation was performed as described in Section 2. Representative gross wound images were acquired at post injection days 0, 4, 8, 12. Skin wound sizes at day 8 were compared with wound size at day 0. n = 5. (B) Tissue slide samples were stained with hematoxylin & eosin. Low & high magnified histological gross images are shown in the left & right panels, respectively. Scale bars, 260 µm (magnification, ×40) & 100 µm (magnification, ×100). n = 5. (C) Representative images of blood vessels in skin wounds on day 12 (top panel). Vessel density was analyzed by using ImageJ program (bottom panel). n = 5. (D-F) Histological tissue samples were immuno-stained with CD31, alpha-SMA, & HNA-specific antibodies & PI for counterstaining. alpha-SMA & HNA-positive cells were visualized by confocal microscopy. The number of CD31 & alpha-SMA-positive cells in high power field (HPF), & the percentage of HNA-positive cells in total cells were analyzed by using Metamorph software. Scale bars, 100 µm (magnification, ×100). n = 5. Data are presented as a mean ± S.E.M. $p < 0.05 versus vehicle group, *p < 0.05 versus UCB-hMSC group given NT siRNA, #p < 0.05 versus UCB-hMSC group given NT siRNA with hypoxia pretreatment, @p < 0.05 versus UCB-hMSC group given BNIP3 siRNA with hypoxia pretreatment. Image collected & cropped by CiteAb from the following publication (https://linkinghub.elsevier.com/retrieve/pii/S2213231717303804), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: BNIP3L Antibody [NBP1-88558] -

Immunocytochemistry/ Immunofluorescence: BNIP3L Antibody [NBP1-88558] - The role of PA in BNIP3 silenced UCB-hMSC survival in the mouse skin wound healing model. (A) Mouse skin wound surgery with UCB-hMSC transplantation was performed as described in Section 2. Representative gross wound images were acquired at post injection days 0, 4, 8, 12. Skin wound sizes at day 8 were compared with wound size at day 0. n = 5. (B) Tissue slide samples were stained with hematoxylin & eosin. Low & high magnified histological gross images are shown in the left & right panels, respectively. Scale bars, 260 µm (magnification, ×40) & 100 µm (magnification, ×100). n = 5. (C) Representative images of blood vessels in skin wounds on day 12 (top panel). Vessel density was analyzed by using ImageJ program (bottom panel). n = 5. (D-F) Histological tissue samples were immuno-stained with CD31, alpha-SMA, & HNA-specific antibodies & PI for counterstaining. alpha-SMA & HNA-positive cells were visualized by confocal microscopy. The number of CD31 & alpha-SMA-positive cells in high power field (HPF), & the percentage of HNA-positive cells in total cells were analyzed by using Metamorph software. Scale bars, 100 µm (magnification, ×100). n = 5. Data are presented as a mean ± S.E.M. $p < 0.05 versus vehicle group, *p < 0.05 versus UCB-hMSC group given NT siRNA, #p < 0.05 versus UCB-hMSC group given NT siRNA with hypoxia pretreatment, @p < 0.05 versus UCB-hMSC group given BNIP3 siRNA with hypoxia pretreatment. Image collected & cropped by CiteAb from the following publication (https://linkinghub.elsevier.com/retrieve/pii/S2213231717303804), licensed under a CC-BY license. Not internally tested by Novus Biologicals.