Proliferation and cell cycle progression of human PASMCs exposed to hypoxia and/or pretreatment with MFG‐E8 silencing. (a–c) Polymerase chain reaction, western blot analysis and ELISA techniques verified the interference efficiency of a small interfering RNA on MFG‐E8; n = 3, *p < .05 compared with the Si NC group. (d) CCK8 proliferation assay of human PASMCs exposed to 2% O2 for 48 hr and/or pretreatment with Si MFG‐E8; n = 3. (e) Representative cell cycle analysis images showed cell cycle progression alterations with hypoxia stimulation for 48 hr and/or Si MFG‐E8 pretreatment, and the average data of G0/G1 phase (f) and S phase (g); n = 3. (h) Representative images of apoptosis analysis showed that neither hypoxia nor MFG‐E8 silencing had a significant effect on apoptosis. (i) Representative Transwell assay images showed cell migration alterations with hypoxia and/or MFG‐E8 silencing and the average number of migration cells (j); n = 3. The results are expressed as the mean ± SEM, *p < 0.05 compared with the 21% O2 + Si NC group, and #
p < .05 compared with the 2% O2+Si NC group. CCK8, cell counting kit‐8; ELISA, enzyme‐linked immunosorbent assay; MFG‐E8, milk fat globule‐EGF factor 8; NC, negative control; PASMC, pulmonary artery smooth muscle cell