Figure 3
From: Pathogenic mutations reveal a role of RECQ4 in mitochondrial RNA:DNA hybrid formation and resolution
RNA:DNA hybrids accumulate on mtDNA in cells expressing P466L mutant RECQ4 proteins. (a) Fold changes in relative mtDNA:gDNA ratio in RECQ4 KD HEK293 cells expressing P466L or ID mutant RECQ4 as compared to cells expressing WT RECQ4. *** indicates p value < 0.005. (b–f) Fold changes in RNA:DNA hybrid frequency at (b) OH (OH), (c) OL (OL), (d) mt-tRNA, (e) COXIII and (f) ND5 gene loci on mtDNA in stable RECQ4 KD HEK293 cells expressing FLAG-RECQ4 P466L or ID mutant constructs as compared to cells expressing WT RECQ4. RNA:DNA hybrid frequency was measured by DRIP as described in Fig. 1f. (h) Fold changes in RNA:DNA hybrid frequency at exon 3 of ACTB (Ex3) in RECQ4 KD HEK293 cells expressing P466L FLAG-RECQ4 constructs compared to RECQ4 KD HEK293 cells expressing the WT proteins were analyzed by DRIP. For all quantitative analyses shown in Fig. 3, each value represents mean ± standard deviation of 3 independent biological experiments, each with 3 triplicate reactions.
