Fig. 3.

Effect of obesity-associated serum factors on survival of B16F10 cells upon DTIC treatment. a Calculation of IC₅₀ value of DTIC in DMEM containing ND or HFD serum. B16F10 cells were chronically grown in medium containing 5% serum collected from ND or HFD C57BL/6J mice for 10 days. These cells were then subjected to DTIC treatment at the indicated concentrations for 48 h. Thereafter, MTT assay was performed. b Rh-123 efflux assay in B16F10 cells cultured in the medium containing serum from experimental mice. Data were acquired on FACS Calibur and analyzed using BD CellQuest Pro software. c, d Long-term survival assay in B16F10 cells cultured in the medium containing serum from experimental mice in the presence of inhibitors of FASN, Cav-1, and P-gp, either alone or together with DTIC (c). Data were quantitated using ImageJ software (d). The data are representative of experiments performed three times; Ceru or C = cerulenin; MCD or M = methyl β-cyclodextrin; Vera or V = verapamil. The results are given as means ± standard deviation; *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 denote significant differences between the groups; NS non-significant