Figure 4
From: Pathogenic mutations reveal a role of RECQ4 in mitochondrial RNA:DNA hybrid formation and resolution
TFAM depletion does not reduce RNA:DNA hybrid accumulation on mtDNA in RECQ4 P466L mutant cells. (a) Fold changes in the mtRNA:mtDNA ratio in RECQ4 KD HEK293 cells expressing FLAG-RECQ4 P466L or ID constructs as compared to cells expressing FLAG-RECQ4 WT proteins. (b) Western blot analysis of TFAM protein expression in WCEs prepared from HEK293 KD cells expressing P466L mutant and treated with control or TFAM shRNA. Tubulin is used as a loading control. FLAG is used to monitor the expression of P466L mutant RECQ4. (c–e) Fold changes in (c) mtRNA:mtDNA ratio, (d) RNA:DNA hybrid frequency normalized to input mtDNA at the indicated loci and (e) relative mtDNA:gDNA ratio in HEK293 KD cells expressing P466L mutant and treated with TFAM shRNA as compared to P466L mutant cells treated with control shRNA as shown in (b). All quantitative analyses shown in Fig. 4 are based on mean ± standard deviation of 3 independent biological experiments, each with 3 triplicate reactions.
