Western Blot: ZEB2 AntibodyBSA Free [NBP1-77179]
Western Blot: ZEB2 Antibody [NBP1-77179] - Lysates from 293T cells transfected Flag-Zeb2 with or without HA-Fbxo45 were immunoprecipitated (IP) with anti-Flag M2 antibody, and then immunoblotted. Total lysates were also used as Input for immunoblotting analysis. Image collected and cropped by CiteAb from the following publication (oncotarget.com/fulltext/2825), licensed under a CC-BY license.
Western Blot: ZEB2 AntibodyBSA Free [NBP1-77179]
Western Blot: ZEB2 Antibody [NBP1-77179] - The protein levels of EMT-TFs were compared between carcinoma cell lines and non- or less-malignant tumor cell lines by using Western-blot analysis. The soft agar colony transformation or migration assay was referred to distinguish malignant degree of the experimental cancer cell lines (data not shown). Malignant degree: prostate cancer cell lines, P69 < M12; breast cancer cell lines, MCF7 < MD-MBA-231; lung cancer cell lines, A549 < H1299; liver cancer cell lines, HepG2 < SMMC7721. 'kD' (kilo-dalton) means the molecular weight of protein. Image collected and cropped by CiteAb from the following publication (oncotarget.com/fulltext/2825), licensed under a CC-BY license.
Immunocytochemistry/ Immunofluorescence: ZEB2 Antibody - BSA Free [NBP1-77179]
Immunocytochemistry/Immunofluorescence: ZEB2 Antibody [NBP1-77179] - Jurkat cells with ZEB2 antibody at 20 ug/mL.
Immunocytochemistry/ Immunofluorescence: ZEB2 Antibody - BSA Free [NBP1-77179]
Immunocytochemistry/Immunofluorescence: ZEB2 Antibody [NBP1-77179] - Analysis of ZEB2 in Jurkat cells with ZEB2 antibody at 20 ug/ml.
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] -
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] - Fbxo45 induces degradation of core EMT-TFsA. Immunoblot for core EMT-TFs in 293T cells transfected Zeb1, Zeb2, Snai1, Snai2 or Twist1 with increasing amount of Fbxo45. B. Immunoblot for detecting endogenous core EMT-TFs in HeLa cells transfected with or without myc-tagged Fbxo45, & harvested after treatment with 20μM of MG132 for 4 hours. C. Immunoblot for endogenous Zeb1, Zeb2, Snai1 & Snai2 in MCF7 cells treated with indicated different concentrations of 17 beta-estrogen for 24 h in the phenol-free medium to induce the expression of endogenous Fbxo45. D. Immunoblot for endogenous core EMT-TFs in HeLa transfected with siRNAs for Fbxo45 or Pam. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25460509), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] -
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] - Fbxo45 induces degradation of core EMT-TFsA. Immunoblot for core EMT-TFs in 293T cells transfected Zeb1, Zeb2, Snai1, Snai2 or Twist1 with increasing amount of Fbxo45. B. Immunoblot for detecting endogenous core EMT-TFs in HeLa cells transfected with or without myc-tagged Fbxo45, & harvested after treatment with 20μM of MG132 for 4 hours. C. Immunoblot for endogenous Zeb1, Zeb2, Snai1 & Snai2 in MCF7 cells treated with indicated different concentrations of 17 beta-estrogen for 24 h in the phenol-free medium to induce the expression of endogenous Fbxo45. D. Immunoblot for endogenous core EMT-TFs in HeLa transfected with siRNAs for Fbxo45 or Pam. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25460509), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] -
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] - Fbxo45 induces degradation of core EMT-TFsA. Immunoblot for core EMT-TFs in 293T cells transfected Zeb1, Zeb2, Snai1, Snai2 or Twist1 with increasing amount of Fbxo45. B. Immunoblot for detecting endogenous core EMT-TFs in HeLa cells transfected with or without myc-tagged Fbxo45, & harvested after treatment with 20μM of MG132 for 4 hours. C. Immunoblot for endogenous Zeb1, Zeb2, Snai1 & Snai2 in MCF7 cells treated with indicated different concentrations of 17 beta-estrogen for 24 h in the phenol-free medium to induce the expression of endogenous Fbxo45. D. Immunoblot for endogenous core EMT-TFs in HeLa transfected with siRNAs for Fbxo45 or Pam. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25460509), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] -
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] - Fbxo45 induces degradation of core EMT-TFsA. Immunoblot for core EMT-TFs in 293T cells transfected Zeb1, Zeb2, Snai1, Snai2 or Twist1 with increasing amount of Fbxo45. B. Immunoblot for detecting endogenous core EMT-TFs in HeLa cells transfected with or without myc-tagged Fbxo45, & harvested after treatment with 20μM of MG132 for 4 hours. C. Immunoblot for endogenous Zeb1, Zeb2, Snai1 & Snai2 in MCF7 cells treated with indicated different concentrations of 17 beta-estrogen for 24 h in the phenol-free medium to induce the expression of endogenous Fbxo45. D. Immunoblot for endogenous core EMT-TFs in HeLa transfected with siRNAs for Fbxo45 or Pam. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25460509), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] -
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] - Functional domains of Fbxo45 for ubiquitination of Zeb2A. Lysates from 293T cells transfected Flag-Zeb2 with or without HA-Fbxo45 were immunoprecipitated (IP) with anti-Flag M2 antibody, & then immunoblotted. Total lysates were also used as Input for immunoblotting analysis. B. Bacterially expressed GST or GST–Fbxo45, -Fbxo45 deltaF-box, -Fbxo45 deltamid, or -Fbxo45 deltaSPRY fusion proteins & Glutathione-Sepharose beads were incubated with lysates of HeLa cells transfected with Zeb2, Snai1 or Snai2, respectively. The proteins associated with GST–tagged Fbxo45 forms, bound on the Glutathione-Sepharose beads were washed five times with the RIPA buffer before immunoblotting. C. Lysates from 293T cells transfected Flag-Zeb2 with HA-Fbxo45, -Fbxo45 deltamid, -Fbxo45 deltaF-box or -Fbxo45 deltaSPRY were immunoprecipitated with anti-Flag or anti-HA antibodies, & immunoprecipitates were resolved by SDS-PAGE for Western-blot analysis. D. U2OS cells transfected with Flag-Zeb2 & HA-Fbxo45, -Fbxo45 deltaF-box or -Fbxo45 deltaSPRY were stained using the primary antibodies of anti-Flag M2 or anti-HA, & the second antibodies of Alexa Fluor 568 anti-mouse or Alexa Fluor 488 anti-Rabbit, respectively. Scale: 25μm. E-I. Zeb2 ubiquitination assays by using IP expriments under different conditions: Flag-Zeb2 with or without HA-Ub (E); Flag-Zeb2 with HA-Ub WT, K48R or K63R (F); Flag-Zeb2 & HA-Ub K48-only with or without myc-Fbox45 (G); Flag-Zeb2 & HA-Ub K48-only with siRNA control, siRNAs for Fbxo45 or siRNAs for Pam (H); Flag-Zeb2 & HA-Ub with HA-Fbxo45, -Fbxo45 deltaF-box or -Fbxo45 deltaSPRY (I). Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25460509), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] -
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] - Fbxo45 is a direct target of miR-27a* that mediates EMT processesA, C. The expression levels of miR-27a* (A) & Fbxo45 mRNA (C) in P69 & M12 were shown according to the data of miRNA transcriptome sequencing & DGE sequencing, respectively [46]. TPM means transcripts per million. B. Real-time PCR in delta deltaCt methods were used to confirm the expression levels of miR-27a* in P69 & M12. One representative experiment out of three was shown. D. Fbxo45 mRNA levels in stable cell lines miR-27a*-overexpressing P69 & miR-27a*-silencing M12 (zipmiR-27a*). P69 or M12 stably infected with the same lentiviral empty vector was used as a control (ctrl). E. Fbxo45 3′-UTR wide-type or mutant form at the position where is complementary to the 5′ seed region of miR-27a* was subcloned into psiCheck2 vector. 293T cells were transfected with psiCheck2 containing WT or mutant Fbxo45 3′-UTR & miR-27a* or non-specific control vector. The Renilla luciferase activity was normalized on the constitutive activity of firefly luciferase. Data are the mean±S.E.M. of three independent experiments. F-G. Immunoblot for Fbxo45, core EMT-TFs (Zeb1, Zeb2, Twist1) & their associated EMT markers (Fibronectin, N-cadherin, Laminin C2, Vimentin & E-cadherin) in stable cell lines P69-miR-27a* or M12-zipmiR-27a* (F); Immunoblot for detection of protein stability of core EMT-TFs under the treatment with 20μM of MG132 for 4 h before harvested (G). GAPDH is for a loading control. H. Immunblot for Fbxo45, N-cadherin, E-cadherin & five core EMT-TFs in stable cell lines PC3-ctrl, PC3-miR-27a* & PC3-zipmiR-27a*. alpha-tubulin is for a loading control. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25460509), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] -
Western Blot: ZEB2 Antibody - BSA Free [NBP1-77179] - Fbxo45 is a direct target of miR-27a* that mediates EMT processesA, C. The expression levels of miR-27a* (A) & Fbxo45 mRNA (C) in P69 & M12 were shown according to the data of miRNA transcriptome sequencing & DGE sequencing, respectively [46]. TPM means transcripts per million. B. Real-time PCR in delta deltaCt methods were used to confirm the expression levels of miR-27a* in P69 & M12. One representative experiment out of three was shown. D. Fbxo45 mRNA levels in stable cell lines miR-27a*-overexpressing P69 & miR-27a*-silencing M12 (zipmiR-27a*). P69 or M12 stably infected with the same lentiviral empty vector was used as a control (ctrl). E. Fbxo45 3′-UTR wide-type or mutant form at the position where is complementary to the 5′ seed region of miR-27a* was subcloned into psiCheck2 vector. 293T cells were transfected with psiCheck2 containing WT or mutant Fbxo45 3′-UTR & miR-27a* or non-specific control vector. The Renilla luciferase activity was normalized on the constitutive activity of firefly luciferase. Data are the mean±S.E.M. of three independent experiments. F-G. Immunoblot for Fbxo45, core EMT-TFs (Zeb1, Zeb2, Twist1) & their associated EMT markers (Fibronectin, N-cadherin, Laminin C2, Vimentin & E-cadherin) in stable cell lines P69-miR-27a* or M12-zipmiR-27a* (F); Immunoblot for detection of protein stability of core EMT-TFs under the treatment with 20μM of MG132 for 4 h before harvested (G). GAPDH is for a loading control. H. Immunblot for Fbxo45, N-cadherin, E-cadherin & five core EMT-TFs in stable cell lines PC3-ctrl, PC3-miR-27a* & PC3-zipmiR-27a*. alpha-tubulin is for a loading control. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25460509), licensed under a CC-BY license. Not internally tested by Novus Biologicals.