Anti hoxa7

Total protein was extracted from hBMSCs transfected with miR-920 mimic, inhibitor, or NC using RIPA buffer and protease inhibitors at a volume ratio of 100:1. Then, the same amount of protein was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Then, the proteins were transferred to cellulose nitrate membranes and washed with TBST with 5% skim milk at room temperature for 1 h. Next, the membranes were incubated with the following primary antibodies: anti-HOXA7 (1:2000, ProteinTech), anti-ALP (1:1000, Abcam), anti-OSX (1:1000, Abcam), anti-p-p38 (1:1000, Cell Signaling Technology), anti-p38 (1:1000, Cell Signaling Technology), anti-p-JNK (1:1000, Cell Signaling Technology), anti-JNK (1:1000, Cell Signaling Technology), and anti-GAPDH (1:3000, ProteinTech) at 4 °C overnight. The membranes were warmed up to room temperature, washed with TBST 3 times, incubated with secondary antibody (1:5000; ProteinTech) for 2 h, washed with TBST 3 times, and then ECL chemiluminescence and development were performed in the dark. Amersham Imager 600 (GE Healthcare) was used to observe the bands on the PVDF membrane.