Fig. 6.

PARP1 regulates miR-204 expression through the IL-6/JAK2/STAT3 pathway. a Rat VSMCs was pre-infected with Scr shRNA or PARP1 shRNA, and then exposed to high Pi for 3 days. The phosphorylation of STAT3 (pSTAT3) and STAT3 were determined by western blot. (n = 5 per group). b–d, VSMCs were infected with Scr shRNA or STAT3 shRNA, along with Ad-Null or Ad-PARP1, and then incubated with osteogenic media for 14 days. MiR-204 expression was determined by qRT-PCR (b). The expression of pSTAT3/STAT3 and Runx2 was analyzed by western blot (c), and the calcium deposition was quantified by the calcium assay (d). (n = 5 per group). e The phosphorylation of JAK2 and SRC, and their total proteins in calcified VSMCs infected with Scr shRNA or PARP1 shRNA were determined by western blot. f The level of IL-6 in the supernatant of calcified VSMCs was detected by ELISA assay. g, h The mRNA (g) and protein (h) levels of IL-6 in the abdominal arteries of CRF rats inoculated with Scr shRNA or PARP1 shRNA were determined by qRT-PCR and western blot. (n = 5 per group). i–k Rat VSMCs were pre-infected with Ad-Null or Ad-PARP1, and then treated with neutralizing antibodies against IL-6 in the osteogenic media for 14 days. The levels of pJAK2/JAK2, pSTAT3/STAT3 and osteogenic markers (OC and Runx2) were determined by western blot (i), and the calcification was detected by Alizarin red S staining (j) and calcium quantification (k). (n = 5 per group). Statistical significance was assessed using one-way ANOVA followed by for multiple comparison and is presented as follows: NS: no significance, **P < 0.01, ^#P < 0.05, ^##P < 0.01, ^&P < 0.05 and ^&&P < 0.01. All values are means ± S.D. Source data are provided as a Source Data file