Fig. 2

Genipin leads to apoptosis through Mcl-1 transcription levels. a AGS cells treated with 75 and 150 μM Genipin for 24 h were harvested for western blotting with the indicated antibodies. b MKN45 (left) and SNU638 (right) cells were treated with 75 and 150 μM Genipin for 24 h. Mcl-1 protein level was detected by immunoblotting. c, d Total mRNA was isolated from AGS cells treated with 75 and 150 μM Genipin for 24 h. mRNA levels of Mcl-1 were measured by qRT-PCR (c) and RT-PCR (d). ** represents a statistically significant difference of P < 0.01. e–g AGS cells were transfected with Mcl-1 siRNA (siMcl-1). Genipin (150 μM) was treated with transfected cells for 24 h. Cell lysates were assessed by qRT-PCR (e), western blotting with cleaved PARP, Mcl-1, and β-actin (f) and flow cytometry (g). h–j AGS cells were transfected with His-Mcl-1 overexpression plasmid. Genipin (150 μM) was treated with transfected cells for 24 h. Protein levels of cleaved PARP, Mcl-1, and β-actin as well as Mcl-1 mRNA levels were assessed by qRT-PCR (h), immunoblotting (i) and flow cytometry (j). ***, P < 0.001