Fig. 7: Exogenous INSIGs and ATF3 blocked miR-9-5p-induced tumor metastasis. | Nature Communications

Fig. 7: Exogenous INSIGs and ATF3 blocked miR-9-5p-induced tumor metastasis.

From: Tumor-derived miR-9-5p-loaded EVs regulate cholesterol homeostasis to promote breast cancer liver metastasis in mice

Fig. 7

a GFP signals in liver and lung from mice receiving tail vein injections of AAV-GFP, AAV-ATF3 and AAV-INSIG1 + INSIG2 virus. Scale bar, 100 μm. b Quantification of gene expression in liver from mice receiving tail vein injections of AAV-GFP, AAV-INSIG1 and AAV-INSIG2 virus. (unpaired two-tailed t test, n = 6 mice per group). c Western blots showing the expression of key proteins related to cholesterol synthesis in liver from mice receiving tail vein injections of AAV-GFP, AAV-INSIG1 and AAV-INSIG2 virus. d Total cholesterol (TC) and triglycerides (TG) in liver in indicated mice (unpaired two-tailed t test, n = 5 mice per group). e Western blots showing protein expression in liver from mice receiving tail vein injections of AAV-GFP and AAV-ATF3 virus. f Representative IHC images showing CH25H staining. Scale bar, 50 μm. g LC-MS/MS analysis of 25-HC level in liver in indicated groups (unpaired two-tailed t test, n = 5 mice for per group). h Body weight was monitored (two-way ANOVA, n = 6 mice per group). i, j Representative bioluminescence images and quantification of mice inoculated with indicated groups (one-way ANOVA, n = 5 mice per group). k Quantification of liver and lung metastatic nodules (unpaired two-tailed t test, n = 6 mice per group). l RNA level of Mki67 in liver and lung in indicated mice (unpaired two-tailed t test, n = 6 mice per group). m Representative IHC images showing Ki67 staining and the overall percentage of Ki67+ tumor cells (one-way ANOVA, n = 5 per group). Scale bar, 50 μm. n Western blots showing proteins in liver from mice receiving tail vein injections of AAV-GFP and AAV-ATF3 virus. o RNA level of M1 and M2 marker gene in liver and lung in indicated mice (unpaired two-tailed t test, n = 6 mice per group). p Purified Kupffer cells were selected with TIM4+, I-A/I-Eint expression and stained and further analyzed for CD163 and CD11c expression. Representative flow cytometry results were shown. q Quantification of the M2/M1 ratio in flow cytometry results (one-way ANOVA, n = 3 independent batches). Data are presented as mean ± s.d. Source data and exact P value are provided as a Source data file.

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