Fig. 3: Reduction of PNPT1 in human renal tubular HK2 cells caused relocation of mt-dsRNAs into cytoplasm.

a Left: immunostaining of PNPT1 (green) and cytosolic dsRNA (J2, red) in HK2 cells treated with or without HG (40 mM, 7d), TGFβ1 (10 ng/mL, 48 h) or LPS (75 μg/mL, 24 h). Nuclei were stained with DAPI (blue). Right: quantification of PNPT1 and cytosolic dsRNA levels in HK2 cells. b Left: immunostaining of PNPT1 (green) and cytosolic dsRNA (J2, red) in HK2 cells treated with or without TGFβ1 (10 ng/mL, 48 h). Prior to TGFβ1 treatment, HK2 cells were infected with or without PNPT1-expressing lentivirus. Right: quantification of PNPT1 levels and cytosolic dsRNA in HK2 cells. c Left: immunostaining of dsRNA (J2, red) and PNPT1 (green) in HK2 cells transfected with PNPT1 siRNA-expressing plasmid (siPNPT1) or control oligonucleotide-expressing plasmid (siCtrl). Right: quantification of PNPT1 and cytosolic dsRNA levels in HK2 cells. d RT–qPCR analysis of cytosolic mt-dsRNAs in HK2 cells transfected with siPNPT1 or siCtrl plasmid. e Cytosolic levels of mt-dsRNAs detected using specific probes against their heavy and light strand in HK2 cells transfected with or without siPNPT1 plasmid. In situ staining of cytosolic mt-ND5 heavy (f) and light strand (g) in HK2 cells transfected with siPNPT1 or siCtrl plasmid. ATP5A1 served as a mitochondria marker. Scale bars, 50 μm. The above experiments were successfully repeated three times. Two-way ANOVA with Sidak’s multiple comparisons test was performed in a–c. Two-tailed unpaired t test was performed for the statistical analyses in f–g, and the results were presented as the mean ± SEM. Source data are provided as a Source Data file.