Fig. 2: SNRNP200 is a crucial regulator of glycolytic TNBCs, promoting tumor proliferation both in vitro and in vivo. | Cell Discovery

Fig. 2: SNRNP200 is a crucial regulator of glycolytic TNBCs, promoting tumor proliferation both in vitro and in vivo.

From: Targeting SNRNP200-induced splicing dysregulation offers an immunotherapy opportunity for glycolytic triple-negative breast cancer

Fig. 2

a Venn diagram depicting the overlap between the MPS2 hub genes and the 42 spliceosome genes whose protein levels were upregulated in TNBC. b MDA-MB-231 cells were cultured in media supplemented with glucose at the indicated concentrations for 16 h. The cell lysates were subjected to immunoblotting. c MDA-MB-231 cells were glucose-starved for 12 h and then stimulated with glucose (25 mM) for the indicated times. The cell lysates were subjected to immunoblotting. d Correlation coefficient heatmap illustrating associations between normalized mRNA expression levels of upregulated U5 snRNP genes in tumors and enrichment scores of metabolic pathways via Spearman’s correlation analysis in the FUSCC-TNBC cohort (*P < 0.05; **P < 0.01). e Diagram depicting the U5 snRNP core components. f Western blot analysis of SNRNP200, EFTUD2, and PRPF8 expression levels in control and SNRNP200-knockdown MDA-MB-231 cells. g MDA-MB-231 cells were immunoprecipitated with an anti-SNRNP200 antibody, with IgG serving as a negative control. The IP data were analyzed by western blot analysis (on the left). qPCR analysis of the U4, U5, and U6 snRNA levels in the input, RNase A−, and RNase A+ groups. The relative U4, U5, and U6 snRNA levels were normalized to that of the input using the 2−Ct method (on the right). qPCR analysis data are presented as the mean ± SEM. h CCK-8 proliferation assay in control and SNRNP200-knockdown MDA-MB-231 cells. i Schematic outline showing the Snrnp200-targeted ASO treatment timeline: 4T1 mouse breast cancer cells were subcutaneously injected into BALB/c mice. When the tumors reached 50–100 mm3, the mice were treated with ASO-Snrnp200 (5 mg/kg subcutaneous injection, twice a week) or PBS (50 µL, subcutaneous injection, twice a week) for 2 weeks (n = 5 mice/group). Tumor growth in different groups. The data are presented as the mean ± SEM. j A representative image of 4T1 tumors illustrating the effect of ASO-Snrnp200 treatment (n = 5 mice/group). k Western blot analysis of mouse Snrnp200 protein expression in tumor tissues from 4T1 cell-derived xenografts. The data are representative of three independent biological replicates. For g–i, the data were compared using Student’s t-test if the data in each group were normally distributed: ***P < 0.001; ns not significant, P > 0.05.

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