Fig. 3

SPP1 promoted SCs proliferation and inhibited the apoptosis through PKCα. a Real-time qPCR and Western blot tested the efficiency of SPP1-siRNAs (si-SPP1) in SCs. b Real-time qPCR and Western blot examined the overexpression of SPP1 (GV146-SPP1) in SCs. c, e EdU analysis of the proliferation rate of SCs treated by si-SPP1, si-SPP1 + PMA, and si-Control (c), and quantification data was shown (e). d, f The apoptosis of SCs was detected by flow cytometry (d), and the apoptosis rate was quantified (f). g, i EdU analyzed the proliferation rate of SCs treated by GV146-SPP1, GV146-SPP1 + Gö6976, and GV146 (g). The proliferation of fold change was quantified (i). h, j Flow cytometry tested the apoptosis rate of SCs. k–m Western blot analysis of PKCα, p-ERK, ERK, Bcl-2, Bax, cleaved Caspase-3, and Caspase-3 protein expression following si-SPP1, si-SPP1 + PMA transfection and treatment in SCs, or following GV146-SPP1, GV146-SPP1  + Gö6976 transfection and treatment in SCs, respectively. Representative blot (k) and quantification of the related expression of proteins (l, m) were presented. Scale bar, 100 μm. Data were shown as mean ± SEM from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001