Fig. 2: Impact of SmD2 deficiency on HCC progression. | Nature Communications

Fig. 2: Impact of SmD2 deficiency on HCC progression.

From: Acetylation-dependent regulation of core spliceosome modulates hepatocellular carcinoma cassette exons and sensitivity to PARP inhibitors

Fig. 2

a Immunoblot demonstrating the effects of doxycycline (DOX) titration on EZ-Tet-pLKO-shSNRPD2 cells, with 72 hours of treatment followed by cell lysis. The experiment was repeated three times with similar results. WCL, Whole Cell Lysate. b Representative images of colony-formation assays of uninduced or DOX-induced shSNRPD2 expression across three HCC cell types. c Quantification of the colony formation assay (n = 3 biologically independent experiments) shown in b for three different cell lines. two-sided t-tests. d Representative confocal images of spheroid formation assays with DOX-induced SNRPD2 knockdown in HCC cell lines, stained with Phalloidin (to stain the cytoskeleton) and DAPI (to stain nuclei). e Quantitative analysis of spheroid diameters (n = 22 biologically independent samples) from the assay in d, showing a significant decrease in spheroid size with SNRPD2 knockdown across all three cell lines. two-sided t-tests. f Tumor growth of inducible shSNRPD2 HCCLM3 cells in male nude mice, with doxycycline (20 mg/kg) orally administered every other day to induce shSNRPD2 expression, beginning on day 7. Two-way ANOVA. g Representative images of tumor volumes from HCCLM3-harboring mice sacrificed on day 29. h Quantitative analysis of tumor (n = 11) volume and weight from (g). two-sided t-tests. i Schematic illustrating transposon- and CRISPR-Cas9-based hydrodynamic injection of vectors into mice via the tail vein to generate liver tumors resembling human HCC. The transposon-based vector overexpressing MYC can express luciferase. j Bioluminescence images of C57BL/6 mice (n = 10) at indicated time points after plasmid injection, indicating tumor burden over time. k Individual bioluminescence trajectories for each mouse over time, comparing non-targeting control to sg-SNRPD2. l Bioluminescence signal at indicated time points post plasmid injection in C57BL/6 mice (n = 10). two-sided t-tests. m Representative images of livers from j. n Kaplan–Meier survival curve of C57BL/6 mice treated as in j. log-rank test. Data are presented as mean values ± SEM. Figure 2i created with BioRender.com, released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license. Source data are provided as a Source Data file.

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