A. Cardiomyocyte-specific overexpression of XBP1s increased mTOR signaling in the heart. XBP1s expression in the heart was turned on for 2 weeks by removal of doxycycline from drinking water. Cardiac tissues were subjected to Western blotting to detect the mTOR signaling. GAPDH was used as loading control.
B. Quantification of A. Normalization was done with phosphorylation levels to respective total proteins. Student’s t test was conducted. N = 4–7.
C. XBP1s overexpression in vitro augmented mTOR activity. NRVMs were infected by adenovirus expressing either GFP or XBP1s. PE treatment was then conducted for 24 hrs.
D. Quantification of C. N = 5 for each group.
E. XBP1s knockdown decreased the mTOR activity. XBP1s expression was reduced by siRNA transfection in NRVMs and PE was incubated for 24 hrs. GAPDH was used as a loading control.
F. Quantification of E. N = 3–6. Two-way ANOVA analysis was conducted, followed by Tukey’s test. *, P<0.05; **, P<0.01; ***, P<0.001.