Figure 2.
PRIMPOL Rescues Nascent Strand Degradation in BRCA1-Deficient Cells
(A–C) PRIMPOL mRNA (A) and protein (B and C) expression 24 h after treatment with 0 or 50 μM cisplatin in UW and UW+BRCA1 cells. (A) Means ± SEM (n = 5) are shown and presented relative to untreated UW+BRCA1 cells. Representative western blot (B) and quantifications (C) from three independent experiments. Statistics: two-way ANOVA followed by Bonferroni test. ns, non-significant, ∗p < 0.05.
(D) Chromatin-bound PRIMPOL in UW cells ± 150 μM cisplatin ± 50 μM mirin ± pre-dose. A representative western blot from three independent experiments is shown. Whole-cell extracts of PRIMPOL KO cells confirm the PRIMPOL antibody specificity.
(E) Expression of PRIMPOL after siRNA (siPRIMPOL) knockdown in UW cells 24 and 48 h after transfection (top). Dot plot and median of IdU tract lengths in siPRIMPOL or siRNA control (siCT) UW cells ± 150 μM cisplatin ± pre-dose (bottom) (n = 3). ns, non-significant, ∗∗p < 0.01, ∗∗∗∗p < 0.0001.
(F) Dot plot and median of IdU tract lengths in siCT or siPRIMPOL UW cells treated with ± 30 J/m2 UVC ± pre-dose (10 J/m2) (n = 3). ns, non-significant, ∗∗∗∗p < 0.0001.
(G) Dot plot and median of CldU/IdU ratios in siCT or siPRIMPOL UW cells treated with 4 mM HU for 2 h ± pre-dose (1 mM HU for 2 h) (n = 3). ns, non-significant, ∗∗∗∗p < 0.0001.
See also Figure S2.