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Fig. 8 | Molecular Cancer

Fig. 8

From: 3-BrPA eliminates human bladder cancer cells with highly oncogenic signatures via engagement of specific death programs and perturbation of multiple signaling and metabolic determinants

Fig. 8

MCT family-member and macropinocytosis activities critically regulate bladder cancer cell susceptibility to 3-BrPA - the dispensable role of MPC components. (a) Representative (three independent experiments) Western blotting profiles of whole-cell protein extracts obtained from RT4 and T24 cells, seeded at ~60 % confluency and treated with the indicated doses of 3-BrPA for 24 h. The proteins examined were MCT1, MCT4, SMCT1, MPC1 and MPC2, while Actin was used as molecule of reference. (b) Representative (three independent experiments) immunofluorescence images of MCT1 expression and localization in RT4 and T24 cells, seeded at ~60 % confluency and exposed to the indicated doses of 3-BrPA for 4 h (also, see Fig. 1c). Scale bars: 3 μm. (c-e) MTT cytotoxicity assays of T24 cells, grown at low (and ~60 %; data not shown) confluency and treated with the indicated doses of 3-BrPA for 24 h, in the absence or presence (pre-incubation for the denoted time) of 1 or 10 μM UK-5099 (c), 0.1 or 1 μM AR-C155858 (d) and 10 or 50 μM EIPA (e) (both low and ~60 % confluency allowed the striking survival of 3-BrPA-treated T24 cells grown in the presence of either AR-C155858 or EIPA, but not UK-5099, inhibitor). Each inhibitor remained in the growth medium with half of its initial respective concentration(s) for 24 h more, post-pre-incubation. Survival rates of each cocktail (3-BrPA plus inhibitor) were normalized according to the respective values of inhibitor only. Stock solutions of all three inhibitors (UK-5099, AR-C155858 and EIPA) (c-e) were prepared in DMSO. Pre-incubation (for 1.5 h) of T24 with 100 μM EIPA proved significantly detrimental for the cells (data not shown). (c-e) Results are reported as mean ± standard deviation of triplicates of three independent experiments. *P < 0.001

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