Fig. 2

Cellular responses to RNA LLPS in CP-treated 4T1 cells. a Signaling pathways (KEGG) related to tumor immunity based on RNA-seq analysis of CP-treated 4T1 cells. b Heatmap of genome-wide RNA-seq profiling of 4T1 cells 24 h after CP treatment. c Venn diagram illustrating the number of tumor immunity-related genes significantly downregulated in CP-treated 4T1 cells. d Copy number of TGFβ1 mRNA transcripts in 4T1 cells after 6 h pf CP treatment and e 30 min of CP treatment. f ELISA measurements of TGFβ1 in CP-treated 4T1 cell culture medium. g Fluorescence in situ hybridization analysis showing TGFβ1 mRNA colocalization with the CPs in 4T1 cells. Scale bar, 5 μm. h Representative double-staining RNA droplet images with lysosomes in 4T1 cells after 4 h of CP treatment and i 15 min of CP treatment. Scale bar, 5 μm. j The signal intensity of PI fluorescence for 4T1 cells after incubation with cationic polymers (Polymer concentration, μg/ml). Data are expressed as the mean ± SEM, and the differences between experimental groups were analyzed by one-way ANOVA with Dunnett’s test (d–f and j). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001