Figure 1.

Dihydroartemisinin upregulates p8, ATF4 and CHOP at transcription level and protein level in HeLa and HCT116 cells. (A). HeLa and HCT116 cells were treated with 20 μM DHA for the indicated times, and total cell extracts were probed with antibodies against p8, ATF4, CHOP and β-actin. (B and C).HeLa and HCT116 cells treated with 20 μM DHA for 9 h, then total RNA was extracted and p8, ATF4 and CHOP mRNA expression was analyzed by RT-PCR, using GAPDH as a control gene. Three independent experiments were performed and the values were expressed as the mean ± SD. (E and H). After transfection with pCCL⁻ p8 plasmid or specifically targeted siRNA (p8 or ATF4) for 24 h, HeLa and HCT116 cells were treated with DHA (20 μM) for 24 h, after which p8, ATF4 and CHOP protein levels were measured by protein gel blot analysis. β-actin was measured as the loading control. (D, F-G) After transfection with specifically targeted p8 siRNA for 24 h, HeLa cells treated with DHA (20 μM) for 9 h, after which total RNA was extracted and p8, ATF4 and CHOP mRNA expression was analyzed by RT-PCR, using GAPDH as a control gene. Three independent experiments were performed and the values were expressed as the mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001.