Fig. 7From: 3-BrPA eliminates human bladder cancer cells with highly oncogenic signatures via engagement of specific death programs and perturbation of multiple signaling and metabolic determinantsCritical regulators of glucose homeostasis are detrimentally targeted after bladder cancer cell exposure to 3-BrPA: drug-induced splicing silencing of GLUT4 RNA. (a-b) Representative (three independent experiments) Western blotting profiles of whole-cell protein extracts obtained from RT4, T24 and T24-X cells, seeded at ~60Â % confluency and exposed to the indicated doses of 3-BrPA for 24Â h. The proteins examined were HK2, GAPDH, p-AS160-Thr642, AS160, Rab10, Tug (a), GLUT1 and GLUT4 (b), while Actin was used as molecule of reference. (c-f) Gene expression profiles, as evidenced through employment of RT-sqPCR protocols (three independent experiments), using total RNA preparations derived from RT4, T24 and T24-X cells, grown at ~60Â % confluency and treated with the indicated doses of 3-BrPA (c-f), Taxol and Doxorubicin (f) for 24Â h (also, see Additional file 5: Figure S5, Additional file 6: Figure S6 and Additional file 7: Figure S7). Besides the oligonucleotide primers able to specifically recognize the GLUT1, GLUT2, GLUT3 (c) and FasL (f) genes, GLUT4-specific primers were used to amplify several exon-exon (e.g. Ex7-Ex8), exon-intron (e.g. Ex4-In4/5) and intra-intronic (e.g. In2/3) segments of the two major RNA splicing variants examined (http://www.ensembl.org/Homo_sapiens) (d-f) (also, see Additional file 5: Figure S5 and Additional file 9: Table S1). GAPDH served as gene of reference. Ex: (single) exon. In: intron (in-between successive exons). Sv001/Sv004: GLUT4 RNA splicing variantsBack to article page