Fig. 4
Expression of mRNA, miRNA and cellular localization of proteins involved in osteogenic differentiation in MC3T3 cells cultured on standard glass slides or on the glass slides covered with HfO2 for 144 h. Relative expression of TGFB, RUNX2, OPN and OCN was determined using RT-qPCR method (a). GAPDH was used as normalization control. Relative expression of osteogenesis-related miRNA (b). U6 was used as endogenous control for miRNA qPCR. Additionally, immunufluorescence for RUNX2 (c), OPN (d) and OPG (e) were performed. Error bars represent the means ± SD. *p < 0.05, **p < 0.01, ***p < 0.001; Student’s t test. Representative images of immunofluorescence staining for RUNX2, OPG and OPN (Z-projects) (green: Atto 488, blue: DAPI) (c). Scale bar is equal 50 µm