Figure 1

iNOS silencing in HT29 cells. (A) Western blot detection of iNOS, eNOS, nNOS and GAPDH in HT29, HT29-dx and HT29 iNOS^- cells. The silencing of iNOS was performed in HT29 cells as reported under the Methods section; then, after a 24 h incubation in the absence (-) or in the presence (+) of 5 μmol/L doxorubicin, cells were lysed and Western blotting was performed as described. The expression of the housekeeping protein GAPDH was measured as equal control loading. The results shown here are representative of three similar experiments. (B). Real Time-PCR for iNOS in HT29, HT29-dx and HT29 iNOS^- cells. After a 24 h incubation in the absence (CTRL) or in the presence of 5 μmol/L doxorubicin (doxo), cells were lysed and Real Time-PCR was performed as described in the Methods section. Data are represented as mean ± SE (n = 3). Vs HT29 CTRL: * p < 0.01; ** p < 0.005. Vs HT29 doxo: ° p < 0.02. (C) HT29 cells were transfected with a non-targeting scrambled shRNA sequence (nt shRNA), then incubated in the absence (-) or in the presence (+) of 5 μmol/L doxorubicin for 24 h and subjected to Western blot for iNOS and GAPDH.