Fig. 2
Overexpression of heat shock factor protein 1 (HSF1) stimulates Prominin-2 expression by activating PROM2 (encoding protein Prominin-2) transcription. A total of 1 × 106 bone mesenchymal stem cells (BMSCs) were used in each group. A BMSCs were incubated with PBS or TBHP (50 μM) for 24 h. Then BMSCs were harvested and PROM2 mRNA levels were determined by RT-PCR. Data were normalized to the levels of GAPDH (n = 5). B Correlation between HSF1 mRNA levels and PROM2 mRNA levels was performed using Pearson correlation analysis. Negative control lentivirus transfected BMSC (LV-NC) or lentiviral vectors carrying HSF1 gene transfected BMSC (LV-HSF1) incubated with PBS or TBHP (50 μM) for 24 h. C BMSCs were harvested and PROM2 mRNA levels were determined by RT-PCR. Data were normalized to the levels of GAPDH (n = 5). D and G BMSCs were harvested, and Prominin-2, cytoplasmic HSF1, and nuclear HSF1 were evaluated by Western Blot (n = 5). Histone-3 was used as a loading control for nuclear fractions. E and F Quantitative analysis of the protein levels of Prominin-2 and cytoplasmic HSF1. H Quantitative analysis of the protein levels of nuclear HSF1. All data are representative of at least five separate experiments. All these experiments were repeated by at least five times. Analysis: unpaired student’s t-test (A), ANOVA followed by a post hoc analysis (C, E, F, and H); **P < 0.01, ***P < 0.001