Fig. 4. Western blot analysis of WM266-4 and M14 melanoma cells confirms that autophagy is induced by 2155-14. (A) LC3 and beclin-1 representative blots and quantification 30 min after addition of 5-50 µM 2155-14 and 2155-18 to WM266-4 cells. No statistical significance between untreated control and test conditions was observed. (B) LC3 and beclin-1 representative blots and quantification 24 h after addition of 5-200 µM 2155-14 to WM266-4 cells. Dose dependent increase of LC3 and beclin-1 was observed in the case of 2155-14, but not 2155-18. (C) LC3 and beclin-1 representative blots and quantification 24 h after addition of 100 µM 2155-14 to M14 cells. UC - untreated control. Rapamycin/chloroquine (Rap/CHQ) mixture used as a control. One-way analysis of variance (ANOVA) was used followed by Dunnett post hoc test. The data shown were the mean ± SD, n=3. ***** - p<0.0001, *** - p<0.001, ** - p<0.01, * - p<0.05.