Chmidoc

The proteins of hPDLCs were harvested with a RIPA lysis buffer (Beyotime, China), and the protein concentration was measured using a BCA protein assay kit (Solarbio, China). Proteins were separated on SDS–PAGE (8–20% gel) and transferred to PVDF membranes. The membranes were blocked for 0.5 h with a blocking reagent, and then were washed and incubated with a primary antibody at 4 °C overnight. After several washes, the membranes were incubated with secondary antibodies (Beyotime, China) at room temperature for 1 h. ECL solution A and solution B (Beyotime, China) were mixed to image the membranes in a darkroom (Bio-Rad ChmiDoc, USA). The primary antibodies were listed in the following manner: GAPDH (ABclonal,1:5000,USA), NFATC1 (ABclonal,1:1000,USA), TIMP-1 (ABclonal,1:1000,USA),TIMP-2 (ABclonal,1:1000,USA), MMP-1 (ABclonal,1:1000,USA),MMP-2 (ABclonal,1:1000,USA), Runx2 (ABclonal,1:1000,USA),MMP-9 (ABclonal,1:1000,USA), COL-I (ABclonal,1:1000,USA), COL-III (ABclonal,1:1000,USA), c-Fos (CST, 1:1000, US), OCN (CST, 1:1000), CTSK (Abcam,1:1000).