Fig. 9

Modulation of HepG2 cell apoptosis induced by etoposide by co-cultured M0, M1 and M2 macrophages. Macrophages were co-cultured in indirect contact with HepG2 cells during 16 h before incubation with or without 50 μM etoposide (+/− e) during 24 h. (a) HepG2 cell proteins were extracted and PARP-1 and caspase-3 protein abundance was assessed by western blotting using specific antibodies. ß-actin was used as loading control. Graphs represent the quantification of cleaved PARP-1 and cleaved caspase-3 abundance normalized by the corresponding ß-actin in three independent experiences. Results are expressed as mean ± 1 S.D. (n = 3). (b) After the incubation with etoposide, caspase-3 and-7 activity was assayed in HepG2 cells by measuring the fluorescence intensity of free AFC released from the cleavage of Ac-DEVD-AFC. Results are expressed in relative caspase-3/-7 activity as mean ± 1 S.D. (n = 3). Statistical analysis was carried out with the one-way ANOVA test followed by a Holm-Sidak post-test. NS: no significantly different from control cells incubated with etoposide; * or **: significantly different from control cells incubated with etoposide respectively with p < 0.05 or 0.01