NUSAP1 Antibody - BSA Free

Western Blot: NUSAP1 Antibody - BSA Free [NBP2-13685] -

NUSAP1 depletion reduces CPT-induced R-loop accumulation and DNA damage and DHX9 depletion increases both. (A) Dot-blot analysis of R-loop levels in LNCaP cells without and with 10 μM Camptothecin (CPT) treatment for 60 min after 48 h post-transfection of siNUSAP1, siDHX9, or both. Serial dilutions of RNA/DNA hybrids (R-loops) and dsDNA extracts were spotted on the membrane for each condition. R-loop formation was increased with CPT treatment. siNUSAP1 alone reduced R-loop levels, while siDHX9 significantly increased R-loops. siDHX9 and siNUSAP1 together did not reduce the accumulation of R-loops compared to siDHX9 alone. Bottom panels anti-DS DNA antibody serves as loading control. (B) S9.6 signal from the dot-blots in the 100 ng nucleic acid samples normalized by dsDNA signal quantified using Image J (n = 3). (C) Immunofluorescent staining using anti-gamma H2AX (red) to assess DNA damage without and with CPT treatment in LNCaP cells. siNUSAP1 decreases gammaH2AX staining while siDHX9 increases staining significantly, which is not affected by siNUSAP1. DAPI (blue). Scale bar = 10 um. (D) gammaH2AX fluorescence intensities quantified by Image J (n = 3). (E) Representative Western blot analysis of NUSAP1, ILF2, DHX9, H2AX, and gammaH2AX protein levels with CPT and the respective siRNA treatments (n = 3). siCtrl, siControl; siN + siD, siNUSAP1 + siDHX9. *, p < 0.05; **, p < 0.01; ****, p < 0.0001; NS, p > 0.05. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37047232), licensed under a CC-BY license. Not internally tested by Novus Biologicals.