Journal: Cancers

Article Title: The Homeobox Transcription Factor NKX3.1 Displays an Oncogenic Role in Castration-Resistant Prostate Cancer Cells

doi: 10.3390/cancers17020306

Figure Lengend Snippet: Expression levels of NKX3.1 vary across PCa cell lines and closely track with AR expression levels. ( A ) Western blot illustrating AR, NKX3.1, and B-actin protein levels in benign prostate cell lines and adenocarcinoma prostate cell lines. ( B ) Western blot illustrating protein levels of AR and NKX3.1 in AR-null cell lines and AR-positive cell lines. ( C ) Co-immunoprecipitation Western blot of LNCaP cells with IgG, AR, FOXA1, and NKX3.1 pulldown. ( D ) Co-immunoprecipitation Western blot of CWR-R1 cells with IgG, AR, FOXA1, and NKX3.1 pulldown. ( E ) Nano-BiT luminescence data depicting binding of large BiT (Lg) and small (Sm) BiT partners on constructs tagged on N- or C-terminally expressed AR, NKX3.1, and/or FOXA1. Significance was determined via one-way ANOVA comparing signal to each individual single BiT. The threshold for statistical significance set as follows: non-significant (ns) p > 0.05, * p < 0.05, *** p < 0.001, **** p < 0.0001.

Article Snippet: Antibodies used: anti-AR (D6F11 XP ® , Cell Signaling Technology, (Danvers, MA, USA)); anti-Beta Actin (AC-15, Sigma-Aldrich); anti-FOXA1 (3A8 PIMA1091, Thermo Fisher); anti-NKX3.1 [D2Y1A XP ® , Cell Signaling Technology, (Danvers, MA, USA)]; anti-Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Cell Signaling, Cat. #2118); and anti-cleaved PARP [Asp214 D64E10 XP ® , Cell Signaling Technology, (Danvers, MA, USA)].

Techniques: Expressing, Western Blot, Immunoprecipitation, Binding Assay, Construct