Extended Data Fig. 6: Validation of siRNA-mediated depletion of PALB2 and RAD52 and specificity of PALB2 and RAD52 immunoprecipitation. | Nature

Extended Data Fig. 6: Validation of siRNA-mediated depletion of PALB2 and RAD52 and specificity of PALB2 and RAD52 immunoprecipitation.

From: BRCA1 and RNAi factors promote repair mediated by small RNAs and PALB2–RAD52

Extended Data Fig. 6

a, Representative immunoblots showing the efficiency of siRNA-mediated depletion of PALB2 and RAD52 in HeLa cells. Representative blot from n = 7 experiments. b, Representative experiment showing γ-H2AX ChIP analyses after RAD52 siRNA-mediated depletion using two separate siRNAs. The histograms depict the mean ± s.d. values of qPCR replicates. Data were analysed by unpaired t-test and compared to the relevant control cells. c, d, co-IP of endogenous BRCA1 in subcellular fractions of HeLa cells (c) and HMECs (d). Representative blot from n = 3 experiments. PALB2 and RAD52 proteins were detected by immunoblotting using the relevant antibodies. Chrom, chromatin; Cyt, cytoplasm; NS, nuclear soluble. e, PALB2 co-IP validation in HeLa cells treated with PALB2 siRNA. Representative blot from n = 2 experiments. f, RAD52 co-IP performed with nuclear extracts from HeLa cells. Representative blot from n = 3 experiments. g, Representative RIP analysis of nuclear PALB2 and RAD52 binding affinity ratio (log2-transformed) between CASP16P sdRNA and its precursor. All immunoblots were developed using the indicated antibodies.

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