rabbit anti orc2  (Bethyl)

Journal: International journal of molecular sciences

Article Title: Extraciliary OFD1 Is Involved in Melanocyte Survival through Cell Adhesion to ECM via Paxillin.

doi: 10.3390/ijms242417528

Figure Lengend Snippet: Figure 5. OFD1 knockdown reduces interactions with the ECM via paxillin. (a,b) Western blot analysis of the relative ratios of paxillin, FAK, and integrin β1 (a) and immunoprecipitation using anti-OFD1 or antipaxillin antibody (b) in cultured melanocytes with or without OFD1 knockdown. (c) Representative immunofluorescent microscopy using anti-OFD1 and antipaxillin antibodies in

Article Snippet: Equal amounts of extracted proteins (20μg) were resolved and transferred to nitrocellulose membranes, which were incubated with antibodies to OFD1 (Novus Biologicals, Centennial, CO, USA), paxillin, FAK, PARP, BAD, BAX, WNT3a (Santa Cruz Biotechnology, Santa Cruz, CA, USA), PTCH1, GLI1, Smo, BCL2, p-BAD, GSK-3β, β-catenin, cleaved caspase-3 (Cell Signaling Technology, Beverly, MA, USA), integrin β1 (Bethyl Laboratories, Montgomery, TX, USA), and β-actin (Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Knockdown, Western Blot, Immunoprecipitation, Cell Culture, Microscopy

Journal: International journal of molecular sciences

Article Title: Extraciliary OFD1 Is Involved in Melanocyte Survival through Cell Adhesion to ECM via Paxillin.

doi: 10.3390/ijms242417528

Figure Lengend Snippet: Figure 7. Downregulation of paxillin by OFD1 knockdown inhibits melanocyte adhesion to the ECM. (a,b) Adhesion assays performed on fibronectin or type IV collagen-coated culture dishes with immunofluorescent microscopy for nuclei (a) or mCherry (b) using cultured melanocytes with control RNA (control sgRNA) or OFD1 knockdown (OFD1 sgRNA) in the presence (paxillin) or absence (mock vector) of mCherry-fused PXN overexpression or nontransfected control (−vector), and cells with mCherry-fused PXN overexpression (bar = 0.05 mm). (c) Western blot analysis of the relative ratios of paxillin, FAK, and integrin β1 in cultured melanocytes with or without OFD1 knockdown in the presence or absence of PXN overexpression. (d,e) Adhesion assay performed on fibronectin- or type IV collagen-coated culture dishes using cultured melanocytes with control sgRNA or PXN knockdown (PXN sgRNA) (d) and cells with IFT88 knockdown (IFT88 sgRNA) (e). β-actin was used as an internal control in the Western blot analysis. The nuclei for immunofluorescence staining were stained with Hoechst 33,258 (bar = 0.05 mm). The data from cultured melanocytes in each graph represent the mean ± SDs of six independent experiments. * p < 0.05 and ** p < 0.01 vs. control, # p < 0.05 and ## p < 0.01 vs. sample without PXN overexpression.

Article Snippet: Equal amounts of extracted proteins (20μg) were resolved and transferred to nitrocellulose membranes, which were incubated with antibodies to OFD1 (Novus Biologicals, Centennial, CO, USA), paxillin, FAK, PARP, BAD, BAX, WNT3a (Santa Cruz Biotechnology, Santa Cruz, CA, USA), PTCH1, GLI1, Smo, BCL2, p-BAD, GSK-3β, β-catenin, cleaved caspase-3 (Cell Signaling Technology, Beverly, MA, USA), integrin β1 (Bethyl Laboratories, Montgomery, TX, USA), and β-actin (Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Knockdown, Microscopy, Cell Culture, Control, Plasmid Preparation, Over Expression, Western Blot, Cell Adhesion Assay, Staining