First, we would like to emphasize that indications of autophagy were diverse and determined from increased cellular granularity, amplified lipidation of LC3 (from LC3-I to LC3-II) detected by Western blot, and elevated levels of LC3 foci observed by fluorescent staining. Nevertheless, to better explain the correlation between autophagy and apoptosis we have performed additional examinations to include more markers and additional time points. As per the reviewer’s request, Beclin-1 levels were increased more than 4-fold relative to control in all treatment groups, while intensity of LC3 staining was increased about 3-fold relative to control in the individual TTFields and sorafenib groups, but elevated only 2-fold in the combination group (Figure 4e).
The in vivo model appears to be performed only time which question the validity of data.