Supplementary Figure 6: Transcriptional manipulation of DNA repair factors using CRISPRi.

a, Stable CRISPRi cells targeting the indicated gene with or without re-expression of a cDNA of the indicated gene were harvested (Fig. 2a,b) and fold depletion of the indicated transcripts over control transcripts (ACTB, GAPDH) was measured by qPCR. The mean ± s.d. was calculated from n = 2 independent experiments. b, Epitope tags reveal robust re-expression of FA proteins. HA blots of parental and cDNA re-expression cell lines are presented along with predicted sizes for re-expressed genes. Data are representative of n = 2 independent experiments. c, Blotting for FANCA confirms knockdown/re-expression data. Top row, FANCA blot of parental (K562) cells, FANCA knockdown CRISPRi cells, and FANCA knockdown CRISPRi cells with re-expressed FANCA. Bottom row, loading controls for each sample. Data are representative of n = 2 independent experiments. d, Blotting for FANCD2 confirms knockdown/re-expression data. Top row, FANCD2 blot of parental (K562) cells, two FANCD2 knockdown cell lines, and FANCD2 knockdown g2 with re-expressed FANCD2. Bottom row, loading controls for each sample. Data are representative of n = 2 independent experiments. e, CD90 abundance validates transcription of cDNA constructs. Re-expression constructs express Thy1.1 co-transcriptionally with the cDNA, which was monitored by live-cell flow cytometry. Data are representative of n = 2 independent experiments.