Figure 4.
Decreased mTOR activity and activation of autophagy after in vitro rapamycin and doxycycline treatment in ZR75.1 cell line. (a) Decreased expressions of mTOR activity-related proteins were assessed by WesTM Simple technique (left panel). β-actin was used as loading control. Densitometric analysis of the studied proteins was presented in the right panel. (b) Time-dependent protein level changes of LC3-B-I/II and p62 detected with Western blot confirmed the autophagy induction. Protein level was normalised to β-actin. Chloroquine (Chl) was the positive control in this experiment. The results LC3-B-II/I ratio and relative expression of p62 based on densitometric analysis of the studied proteins were presented in the middle and lower panels. Dashed lines show the obtained level of control values. Data represent mean ± SD. (Co—control; R—rapamycin, 50 ng/mL; D—doxycycline, 10 µM; R+D—rapamycin + doxycycline combination; Chl—chloroquine, 50 µM).