Figure 6.

AtRNH1C Coimmunoprecipitates with AtGyrases in a DNA- and RNA-Dependent Manner.

(A) AtRNH1C coimmunoprecipitates with AtGyrA. After immunoprecipitation, anti-FLAG and anti-AtGyrA antibodies were used to detect AtRNH1C-3XFLAG and AtGyrA proteins, respectively. T, total protein; S, supernatant; B, beads with copurified proteins.

(B) to (D) AtRNH1C interacts with AtGyrases mediated by DNA/RNA. After immunoprecipitation with anti-FLAG M2 beads, anti-AtGyrA antibody was used to detect AtGyrA proteins. RT-PCR and digestion control were performed by amplifying a 16S rRNA fragment from cDNA reverse transcribed from RNA or DNA isolated from beads with or without nucleases treatment. Primers are listed in Supplemental Table 2. T, total protein; S, supernatant; B, immunoprecipitated beads with copurified proteins; BI, immunoprecipitated beads with DNase I treatment; BA, immunoprecipitated beads with RNase A treatment; BIA, immunoprecipitated beads with DNase I and RNase A treatment.