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).
LC3 is increased in all the groups except in control tissue, however, cell death was increased only in combined group. How can the authors correlate autophagy with apoptosis? It is not so trustworthy that the only expression of LC3B indicate the treated condition have increased autophagy in the tumor tissue. Other important autophagy and degradation markers like Beclin1 and P62 need to be shown to reflect the regulatory mechanism of TTFields, as well as for the combined treatment with Sorafenib.