psa a67 b e3 ab  (Santa Cruz Biotechnology)

Journal: Translational Cancer Research

Article Title: ZBTB7B modulates the androgen receptor as an upstream regulator via colocalization and direct binding in LNCaP prostate cancer cells

doi: 10.21037/tcr-2025-1365

Figure Lengend Snippet: Differentially expressed profile in ZBTB7B depleted LNCaP cells. (A) ZBTB7B expression levels of ZBTB7B in ZBTB7B depleted LNCaP cells were assessed by Western blotting. (B) AR and PSA expression in ZBTB7B depleted LNCaP cells compared to untreated control by scatter plot. (C) GO analysis for associated signaling pathways by ZBTB7B depletion compared to untreated control. (D) Upregulated and downregulated genes in LNCaP sicon and LNCaP siZBTB7B cells by mRNA-Seq-based analysis. Red and green colors in the heat map indicate increased and decreased expression levels, respectively. (E) KEGG pathway of ZBTB7B repressed signaling pathway (green) and induced signaling pathway (red). AR, androgen receptor; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; PSA, prostate-specific antigen.

Article Snippet: Membranes were blocked with 5% skim milk or BSA and incubated overnight at 4 °C with primary antibodies against AR, PSA, ZBTB7B, Flag, His, HA, Lamin B (Santa Cruz Biotechnology, Dallas, TX, USA), or β-actin (Sigma).

Techniques: Expressing, Western Blot, Control, Protein-Protein interactions

Journal: Translational Cancer Research

Article Title: ZBTB7B modulates the androgen receptor as an upstream regulator via colocalization and direct binding in LNCaP prostate cancer cells

doi: 10.21037/tcr-2025-1365

Figure Lengend Snippet: ZBTB7B regulates AR as an upstream regulator via direct binding. (A) Correlation coefficient between ZBTB7B and AR mRNA level in prostate cancer. (B) Effects of ZBTB7B depletion on AR mRNA level in LNCaP cells. (C) Effect of AR depletion on ZBTB7B mRNA level in LNCaP cells. (D) Effects of ZBTB7B depletion on AR at protein levels in LNCaP cells. (E) Effects of ZBTB7B depletion on PSA secretion in LNCaP cells, as measured by ELISA. (F) AR depletion does not affect ZBTB7B at protein level in LNCaP cells. (G) Effects of ZBTB7B depletion on AR mRNA stability in LNCaP cells in the presence or absence of cycloheximide. (H) Schematic representation of AR domains. (I) The NTD domain of AR is requisite for binding with ZBTB7B in HEK293 cells. Cells were co-transfected with HA-HA-AR-NTD, HA-HA-AR-NTD-DBD, HA-HA-AR-DBD-LBD and/or ZBTB7B plasmids and then were subjected to immunoprecipitation with anti-HA antibody and HA magnetic beads and were immunoblotted with HA and Flag. **, P<0.01; ***, P<0.001. AR, androgen receptor; AR-FL, AR-full length; CHX, cycloheximide; DBD, DNA-binding domain; LBD, ligand-binding domain; NTD, N-terminal domain; PSA, prostate-specific antigen.

Article Snippet: Membranes were blocked with 5% skim milk or BSA and incubated overnight at 4 °C with primary antibodies against AR, PSA, ZBTB7B, Flag, His, HA, Lamin B (Santa Cruz Biotechnology, Dallas, TX, USA), or β-actin (Sigma).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Transfection, Immunoprecipitation, Magnetic Beads, Ligand Binding Assay

Journal: Translational Cancer Research

Article Title: ZBTB7B modulates the androgen receptor as an upstream regulator via colocalization and direct binding in LNCaP prostate cancer cells

doi: 10.21037/tcr-2025-1365

Figure Lengend Snippet: Effects of finasteride or DHT on the expression of ZBTB7B in LNCaP cells. (A) Effects of finasteride on the expression of ZBTB7B, AR and PSA in LNCaP cells. (B) Effect of DHT on the expression of ZBTB7B, AR, and PSA in LNCaP cells. (C) Effects of ZBTB7B depletion on the expression of AR and PSA in LNCaP cells exposed to DHT and/or finasteride. (D) Binding between ZBTB7B and AR in the presence or absence of DHT. (E) The colocalization between ZBTB7B and AR in LNCaP cells (immunofluorescence staining, scale bar: 40 μm). (F) Effects of ZBTB7B depletion on AR and PSA in the cytosolic and nuclear fractions of LNCaP cells. (G) Effects of ZBTB7B depletion on AR in LNCaP cells with or without MG132 treatment. (H) Effects of ZBTB7B depletion on AR degradation in LNCaP cells as determined by ubiquitination assay. AR, androgen receptor; DHT, dihydrotestosterone; PSA, prostate-specific antigen.

Article Snippet: Membranes were blocked with 5% skim milk or BSA and incubated overnight at 4 °C with primary antibodies against AR, PSA, ZBTB7B, Flag, His, HA, Lamin B (Santa Cruz Biotechnology, Dallas, TX, USA), or β-actin (Sigma).

Techniques: Expressing, Binding Assay, Immunofluorescence, Staining, Ubiquitin Proteomics