Fig 1: Expression of prolyl 4-hydroxylase subunit alpha P4ha gene and hydroxyproline protein level increase with vesicle trafficking gene knockdown in MC3T3-E1 osteogenic differentiation. (A) Ten days after osteogenic differentiation, relative P4ha1 and P4ha2 mRNA expression in control and trafficking gene knockdown MC3T3-E1 cell differentiation were analyzed by qRT-PCR. Y axes show relative P4ha1 and P4ha2 mRNA expression in trafficking gene knockdown normalized to siControl. All data represent with box-whiskers plot (N = 6–9; multiple t test; *P < 0.05; **P < 0.01; #P < 0.001; relative to siControl). (B) Ten days after osteogenic differentiation, Western-blot analysis shows Hydroxyproline and GAPDH protein expression in control and trafficking gene knockdown MC3T3-E1 cell differentiation. Full gel blots are included in Supplementary Fig. S11. All data represent with scatter plots with mean ± SD (N = 3–6; multiple t test; *P < 0.05; **P < 0.01; #P < 0.001; relative to siControl).