Fig. 2. MKK3 depletion affects cell growth via induction of autophagy in a subset of CRC lines.

a Relative cell number of sh/scr and sh/MKK3 sublines after 144 h doxycycline (1 μg/ml) treatment were determined by trypan blue exclusion assay and numbers of viable cells reported as mean and S.D. of three independent experiments. Significance was analyzed using unpaired t-test. Results from each sh/MKK3 subline were normalized to those of relative control (sh/scr subline) set to 1.0. b Western blot analyses were performed on protein lysates derived from sh/scr and sh/MKK3 sublines after 144 h doxycycline (1 μg/ml) treatment. Blots were incubated with indicated antibodies. More relevant bands from the same filter at same exposure length are reported. Representative results of three independent experiments are reported. c LC3II/LC3I ratio and data are reported as mean and S.D. of three independent experiments. Densitometry was performed with ImageJ software and quantified with respect to controls (sh-scr) set to 1.0. Significance was analyzed using unpaired t-test. *p < 0.05, ***p < 0.001, ****p < 0.0001, n.s.: not significant