WDR34 Antibody

Knockdown Validated: WDR34 Antibody [NBP1-88805]

Knockdown Validated: WDR34 Antibody [NBP1-88805] - The stabilities of WDR34 and WDR60 are interdependent. Immunoblotting confirmed the efficacy of WDR34 and WDR60 siRNAs. Immunoblotting for WDR34 following suppression of WDR60 and vice versa was also used to test the interdependency of these subunits. Lamin A/C is included as a negative control, GAPDH as a loading control. Molecular-mass markers are indicated (kDa). Image collected and cropped by CiteAb from the following publication (https://jcs.biologists.org/cgi/doi/10.1242/jcs.159038), licensed under a CC-BY license.

Knockout Validated: WDR34 Antibody [NBP1-88805]

Knockout Validated: WDR34 Antibody [NBP1-88805] - HA-WDR34 (NBP1-88805) and HA-WDR60 (NBP1-90437) expression in WT and KO cells. (A) Pull down of HA-WDR60 in WT and WDR34 KO cells. HA-WDR60 pulls down WDR34 in WT but not in KO cells (B) Pull down of HA-WDR34 in WT and WDR60 KO cells. HA-WDR34 pulls down WDR60 in WT but not in KO cells. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30320547/) licensed under a CC-BY license.

Western Blot: WDR34 Antibody [NBP1-88805] -

Western Blot: WDR34 Antibody [NBP1-88805] - Immunoprecipitation from serum-starved RPE1 cells stably expressing either mGFP or mGFP–WDR34. GFP-traps of cells expressing mGFP alone or mGFP–WDR34 were separated by SDS-PAGE & immunoblotted to detect (A) WDR34, (B) WDR60, (C) IC74 (DYNC1I2), (D) GAPDH, (E) p150Glued, (F) p50dynamitin, (G) LIS1, (H) Tctex-1, (I) NudCD3 or (J) TCTEX1D2, as indicated. Molecular-mass markers are shown (kDa). In each case, the lanes show the input (‘I’), the unbound fraction (‘U’) & the bound fraction (‘B’), as indicated. The asterisk in J indicates non-specific bands. Image collected & cropped by CiteAb from the following publication (https://journals.biologists.com/jcs/article/doi/10.1242/jcs.159038/259056/Subunit-composition-of-the-human-cytoplasmic), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: WDR34 Antibody [NBP1-88805] -

Western Blot: WDR34 Antibody [NBP1-88805] - (A) RNAi of WDR34 & WDR60 validated by immunoblotting with GAPODH as a loading control.(B) Immunofluorescence of TMEM67 & RPGRIP1L in WDR34 & WDR60 depleted cells. TMEM67 is seen at the base & within the cilium proximal to this in both control & depleted cells, RPGRIP1L is more tightly restricted to the base of the cilium in all cases. These are representative images, we have analysed multiple fields of view in each case using z-stacks & cannot identify any difference. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30320547), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: WDR34 Antibody [NBP1-88805] -

Western Blot: WDR34 Antibody [NBP1-88805] - CRISPR control cells lines show no defect in ciliogenesis.(A & B) Immunoblotting for WDR34 & WDR60 in WT & KO cells. (A) A 52 kDa band, corresponding to WDR34 is lost in WDR34 KO cells lysate. (B) A WDR60 band of 123 kDa is lost in the WDR60 KO cells lysate. (C) WDR34 KO CTRL & WDR60 KO CTRL cells stained with Arl13b (green) & AcTub (red). (D) WDR60 KO CTRL cells stained with IFT88 (green). (Di) IFT88 intensity quantification in WT & WDR60 KO CTRL cells (n = 3, 98 WT, 102 WDR60 KO CTRL, & 122 WDR60 KO cells quantified). Mann-Whitney test was used, p-value: *=<0.05 as above. Scale bars 5 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30320547), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: WDR34 Antibody [NBP1-88805] -

Western Blot: WDR34 Antibody [NBP1-88805] - HA-WDR34 & HA-WDR60 expression in WT & KO cells.(A) Pull down of HA-WDR60 in WT & WDR34 KO cells. HA-WDR60 pulls down WDR34 in WT but not in KO cells (B) Pull down of HA-WDR34 in WT & WDR60 KO cells. HA-WDR34 pulls down WDR60 in WT but not in KO cells. (C) Immunoprecipitation of HA-tagged GFP, WDR60, & WDR34 followed by immunoblot for LIC3. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30320547), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: WDR34 Antibody [NBP1-88805] -

Western Blot: WDR34 Antibody [NBP1-88805] - HA-WDR34 & HA-WDR60 expression in WT & KO cells.(A) Pull down of HA-WDR60 in WT & WDR34 KO cells. HA-WDR60 pulls down WDR34 in WT but not in KO cells (B) Pull down of HA-WDR34 in WT & WDR60 KO cells. HA-WDR34 pulls down WDR60 in WT but not in KO cells. (C) Immunoprecipitation of HA-tagged GFP, WDR60, & WDR34 followed by immunoblot for LIC3. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30320547), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: WDR34 Antibody [NBP1-88805] -

Immunocytochemistry/ Immunofluorescence: WDR34 Antibody [NBP1-88805] - Dynein-2 assembly in primary cilium.(A) Immunoblotting for WDR60 & WDR34 in WT, WDR34 KO#1 & WDR60 KO cells. Arrows indicate WDR34 & WDR60 proteins. (B) LIC3/DYNC2LI1 localization in the cilia of WT, WDR34 KO#1 & WDR60 KO cells. (C) DHC2/DYNC2H1 localization at the ciliary base in WT & KO cells. (Ci) Intensity quantification shows a reduction of DHC2/DYNC2H1 at the ciliary base in WDR34 KO#1 cells (n = 3, 120 WT, 106 WDR60 KO, & 71 WDR34 KO #1 cells quantified). (D) TCTEX1/DYNLT1 localizes at the ciliary base in WT & KO cells. (Di) Intensity quantification of TCTEX1/DYNLT1 at the ciliary base (n = 3 115 WT, 85 WDR60 KO, & 50 WDR34 KO#1 cells quantified). Mann-Whitney test, p-value: ****=<0.0001. Scale bars 5 μm. Arrows point to the ciliary base.Overexpression of WDR34 cannot rescue WDR60 KO phenotype & vice versa.(A) HA-WDR60 localization in WT & WDR34 KO cells. (Ai) Intensity quantification of HA-WDR60 in primary cilia (n = 3, 50 WT & 50 WDR34 KO cells quantified). Mann-Whitney test was used, p-value: ****=<0.0001. (B) HA-WDR34 localization in WT & WDR60 cells. For HA immunolabeling in Fig. A & B cells were treated with cytoskeletal buffer as described in methods. (C) Arl13b staining of WDR34 KO cells expressing HA-WDR60. HA labeling in green shows stable expression of HA-WDR60. (D) IFT88 localizes predominantly at the ciliary base in WT cells whereas WDR60 KO cells stably expressing HA-WDR34 accumulate IFT88 at the ciliary tip, similar to untransfected WDR60 KO cells. Scale bars 5 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30320547), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: WDR34 Antibody [NBP1-88805] -

Western Blot: WDR34 Antibody [NBP1-88805] - Dynein-2 assembly in primary cilium.(A) Immunoblotting for WDR60 & WDR34 in WT, WDR34 KO#1 & WDR60 KO cells. Arrows indicate WDR34 & WDR60 proteins. (B) LIC3/DYNC2LI1 localization in the cilia of WT, WDR34 KO#1 & WDR60 KO cells. (C) DHC2/DYNC2H1 localization at the ciliary base in WT & KO cells. (Ci) Intensity quantification shows a reduction of DHC2/DYNC2H1 at the ciliary base in WDR34 KO#1 cells (n = 3, 120 WT, 106 WDR60 KO, & 71 WDR34 KO #1 cells quantified). (D) TCTEX1/DYNLT1 localizes at the ciliary base in WT & KO cells. (Di) Intensity quantification of TCTEX1/DYNLT1 at the ciliary base (n = 3 115 WT, 85 WDR60 KO, & 50 WDR34 KO#1 cells quantified). Mann-Whitney test, p-value: ****=<0.0001. Scale bars 5 μm. Arrows point to the ciliary base.Overexpression of WDR34 cannot rescue WDR60 KO phenotype & vice versa.(A) HA-WDR60 localization in WT & WDR34 KO cells. (Ai) Intensity quantification of HA-WDR60 in primary cilia (n = 3, 50 WT & 50 WDR34 KO cells quantified). Mann-Whitney test was used, p-value: ****=<0.0001. (B) HA-WDR34 localization in WT & WDR60 cells. For HA immunolabeling in Fig. A & B cells were treated with cytoskeletal buffer as described in methods. (C) Arl13b staining of WDR34 KO cells expressing HA-WDR60. HA labeling in green shows stable expression of HA-WDR60. (D) IFT88 localizes predominantly at the ciliary base in WT cells whereas WDR60 KO cells stably expressing HA-WDR34 accumulate IFT88 at the ciliary tip, similar to untransfected WDR60 KO cells. Scale bars 5 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30320547), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: WDR34 Antibody [NBP1-88805] -

Immunocytochemistry/ Immunofluorescence: WDR34 Antibody [NBP1-88805] - Dynein-2 assembly in primary cilium.(A) Immunoblotting for WDR60 & WDR34 in WT, WDR34 KO#1 & WDR60 KO cells. Arrows indicate WDR34 & WDR60 proteins. (B) LIC3/DYNC2LI1 localization in the cilia of WT, WDR34 KO#1 & WDR60 KO cells. (C) DHC2/DYNC2H1 localization at the ciliary base in WT & KO cells. (Ci) Intensity quantification shows a reduction of DHC2/DYNC2H1 at the ciliary base in WDR34 KO#1 cells (n = 3, 120 WT, 106 WDR60 KO, & 71 WDR34 KO #1 cells quantified). (D) TCTEX1/DYNLT1 localizes at the ciliary base in WT & KO cells. (Di) Intensity quantification of TCTEX1/DYNLT1 at the ciliary base (n = 3 115 WT, 85 WDR60 KO, & 50 WDR34 KO#1 cells quantified). Mann-Whitney test, p-value: ****=<0.0001. Scale bars 5 μm. Arrows point to the ciliary base.Overexpression of WDR34 cannot rescue WDR60 KO phenotype & vice versa.(A) HA-WDR60 localization in WT & WDR34 KO cells. (Ai) Intensity quantification of HA-WDR60 in primary cilia (n = 3, 50 WT & 50 WDR34 KO cells quantified). Mann-Whitney test was used, p-value: ****=<0.0001. (B) HA-WDR34 localization in WT & WDR60 cells. For HA immunolabeling in Fig. A & B cells were treated with cytoskeletal buffer as described in methods. (C) Arl13b staining of WDR34 KO cells expressing HA-WDR60. HA labeling in green shows stable expression of HA-WDR60. (D) IFT88 localizes predominantly at the ciliary base in WT cells whereas WDR60 KO cells stably expressing HA-WDR34 accumulate IFT88 at the ciliary tip, similar to untransfected WDR60 KO cells. Scale bars 5 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30320547), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: WDR34 Antibody [NBP1-88805] -

Immunocytochemistry/ Immunofluorescence: WDR34 Antibody [NBP1-88805] - Dynein-2 assembly in primary cilium.(A) Immunoblotting for WDR60 & WDR34 in WT, WDR34 KO#1 & WDR60 KO cells. Arrows indicate WDR34 & WDR60 proteins. (B) LIC3/DYNC2LI1 localization in the cilia of WT, WDR34 KO#1 & WDR60 KO cells. (C) DHC2/DYNC2H1 localization at the ciliary base in WT & KO cells. (Ci) Intensity quantification shows a reduction of DHC2/DYNC2H1 at the ciliary base in WDR34 KO#1 cells (n = 3, 120 WT, 106 WDR60 KO, & 71 WDR34 KO #1 cells quantified). (D) TCTEX1/DYNLT1 localizes at the ciliary base in WT & KO cells. (Di) Intensity quantification of TCTEX1/DYNLT1 at the ciliary base (n = 3 115 WT, 85 WDR60 KO, & 50 WDR34 KO#1 cells quantified). Mann-Whitney test, p-value: ****=<0.0001. Scale bars 5 μm. Arrows point to the ciliary base.Overexpression of WDR34 cannot rescue WDR60 KO phenotype & vice versa.(A) HA-WDR60 localization in WT & WDR34 KO cells. (Ai) Intensity quantification of HA-WDR60 in primary cilia (n = 3, 50 WT & 50 WDR34 KO cells quantified). Mann-Whitney test was used, p-value: ****=<0.0001. (B) HA-WDR34 localization in WT & WDR60 cells. For HA immunolabeling in Fig. A & B cells were treated with cytoskeletal buffer as described in methods. (C) Arl13b staining of WDR34 KO cells expressing HA-WDR60. HA labeling in green shows stable expression of HA-WDR60. (D) IFT88 localizes predominantly at the ciliary base in WT cells whereas WDR60 KO cells stably expressing HA-WDR34 accumulate IFT88 at the ciliary tip, similar to untransfected WDR60 KO cells. Scale bars 5 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30320547), licensed under a CC-BY license. Not internally tested by Novus Biologicals.