Fig. 4

Autophagy induction by FUra and Oxa is dependent of RAC3 levels: Cells lines were loaded in 24 well plates with slices and after 24 h cells were stimulated with 3.5 μM FUra or 0.4 μM Oxa. a, c and f Autophagy was determined by staining with monodansylcadaverine (MDC) after 1 or 6 h post-treatment. b, d, g Diagram bars correspond to percentage of MDC positive cells per field (at least 10 fields per sample). Statistical analysis ANOVA and Tukey post-test n = 3 were performed, b *p < 0.01 3.5 μM FUra at 6 h HT-29 respect to HT-29 basal, **p < 0.001 LoVo basal respect to HT-29 and HCT 116 basal, LoVo treated at each time respect to LoVo basal, HT-29 0.4 μM Oxa at 6 h respect to HT-29 basal and HCT 116 treated for 6 h with FUra or Oxa respect to HCT 116 basal. d *p < 0.001 3.5 μM FUra and 0.4 μM Oxa at 6 h HCT 116 control respect to HCT 116 control basal, **p < 0.001 HCT 116 shRAC3 treated respect to shRAC3 basal, ***p < 0.001 shRAC3 respect to control and g *p < 0.01 3.5 μM FUra LoVo control respect to Lovo control basal and LoVo RAC3 treated with 0.4 μM Oxa for 1 h respect to LoVo RAC3 basal, **p < 0.001 LoVo RAC3 basal respect LoVo control basal, 3.5 μM FUra at any time LoVo control respect LoVo control basal and 0.4 μM Oxa LoVo RAC3 at 6 h respect to LoVo RAC3 basal. e, h LC3-II/I levels were determined by Western blot. RDU correspond to the average of densitometry units, respect to Tubulin expression