Figure 4.

HDAC1 deacetylates LSD1 and thus mediates autophagy in high phosphate‐induced VC models. A, LSD1 expression in VSMCs without treatment or treated with high Pi detected using RT‐qPCR. B, The enrichment of HDAC1 in the promoter region of LSD1 measured using the ChIP assay. C, The enrichment of H3K9ac in the promoter region of LSD1 measured using ChIP assay. D, LSD1 expression in cells treated with oe‐HDAC1 or SAHA (inhibitor of HDAC1) detected using RT‐qPCR. E, Western blot analysis of LSD1 and HDAC1 protein in cells. F, Western blot analysis of LC3 II and p62 proteins in cells. G, The formation of autophagosomes observed under an electron microscope (20 000×). BUN, blood urea nitrogen; CRF, chronic renal failure; GAPDH, glyceraldehyde‐3‐phosphate dehydrogenase; HDAC1, histone deacetylase 1; HE, haematoxylin‐eosin; NC, negative control; oe, overexpression; Pi, inorganic phosphate; RT‐qPCR, reverse transcription‐quantitative polymerase chain reaction; Runx2, Runt‐related transcription factor 2; SCr, serum creatinine; U‐pro, urine protein; VC, vascular calcification; VPA, valproic acid; VSMCs, vascular smooth muscle cells; α‐SMA, α‐smooth muscle actin. *P < .05 indicates significant difference. Data (mean ± SD) between two groups were analysed using unpaired t test, while data among multiple groups were assessed using one‐way ANOVA with Tukey's post hoc test. The experiment was run in triplicate