Fig. 5

Autophagy induction by FUra and OXA induces a decrease in protein acetylation in HCT 116 cells: a, c and e Cells were pre-treated with a deacetylase inhibitor TSA (0.4 μM) for 30 min and then, stimulated with FUra (3.5 μM) or Oxa (0.4 μM) for 6 or 24 h. The arrows show the positive acetylation detected by immunofluorescence using an anti-Lysine acetylated antibody and an antibody anti-mouse coupled to Rodamine. A cell detail of the 10 × magnification is shown in the square. b, d and f Diagram bars correspond to percentage of acetylated protein per field (at least 10 fields per sample). Statistical analysis ANOVA and Tukey post-test n = 3 were performed. b *p < 0.001 LoVo TSA respect to HT-29 and HCT 116 TSA, HCT 116 treated respect to HCT 116 TSA, d *p < 0.01 HCT 116 control TSA plus 3.5 μM FUra at 24 h respect to HCT 116 control TSA, **p < 0.001 HCT 116 control TSA plus FUra or Oxa at 6 h respect HCT 116 control TSA and HCT 116 shRAC3 TSA respect HCT 116 control TSA and f *p < 0.001 LoVo RAC3 with TSA respect to LoVo control TSA. g Expression levels of CD39 in CRC cell lines. The diagram bars show the average ± S.D. of CD39 expression normalized to GAPDH from three independent experiments, *p < 0.001 HCT 116 shRAC3 and LoVo control respect to HCT 116 control and Lovo RAC3 respect to LoVo control. h The diagram bars show the average ± S.D. of CD39 expression normalized to GAPDH from three independent experiments, *p < 0.001 HCT 116 shRAC3 basal respect to HCT 116 control basal and **p < 0.01 cells treated with 0.4 μM Oxa respect to HCT 116 control basal or HCT 116 shRAC3 basal