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. 2014 May 14;9(5):e92786. doi: 10.1371/journal.pone.0092786

Figure 2. Enhanced expression of GM3 synthase and its product GM3 in HCT116 cells after CDDP treatment.

Figure 2

(A) Total RNA from HCT116 cells was isolated following treatment with the indicated concentrations of CDDP treatment for 12 h and GM3 synthase mRNA was detected by RT-PCR. (B) HCT116 cells were transiently transfected with GM3 synthase gene promoter (pGL3-1600) and CREB mutation of GM3 synthase promoter (pGL3-1600 CREB Mu), and then cultured with or without CDDP for 12 h. Luciferase activity was determined from cell lysates as described in the Materials and Methods. The results shown are means ± SD of three independent experiments with triplicate measurement. ***P<0.001 vs. the CDDP-treated control. (C) HCT116 cells were cultured and then incubated with indicated concentration of CDDP treatment for 12 h. Immunoreactivity against ganglioside GM3 was detected using a fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgM without or with the M2590 antibody. (D) The gangliosides isolated from the HCT116 cells treated with CDDP (0, 10, and 30 µg/mL) were analyzed by HPTLC.