Fig. 2: Functional identification of ARID1A as a robust antiviral mediator. | Nature Communications

Fig. 2: Functional identification of ARID1A as a robust antiviral mediator.

From: Virally programmed extracellular vesicles sensitize cancer cells to oncolytic virus and small molecule therapy

Fig. 2

a Fold change in expression of amiR-4 target genes determined by RT-qPCR in MIA PaCa-2 cells infected for 36 h with VSV∆51-amiR-NTC or VSV∆51-amiR-4 (MOI 1) compared to virus control and normalised to loading control (Rplp0). Two-tailed unpaired t test, *P = 0.0107, ****P < 0.0001. Shown are mean values ± SEM of five biological replicates. b Immunoblot analysis showing protein expression levels of amiR-4-predicted targets, GAPDH (loading control), and VSV∆51 proteins in PANC1 cells, subjected or not to VSV∆51-amiR-NTC or VSV∆51-amiR-4 infection for 24 h and 48 h (n = 3). c–h Crystal violet staining pictures and cell viability measured by alamarBlue® Assay of 786-0 wild-type (WT) or ARID1A-knockout (ARID1A−/−) cells mock-infected or infected at multiple MOIs for 48 h with multiple oncolytic virus platforms. Crystal violet staining pictures represent (n = 3) cells mock-infected or infected with VSV∆51-GFP at MOI 1 (c), VSV∆51-amiR-4 at MOI 1 (d), VV TK− VGF− at MOI 1 (e), oHSV-1 GFP at MOI 0.1 (f), Reovirus at MOI 1 (g), or SV-GFP at MOI 1 (h). Two-way ANOVA Tukey’s multiple comparison test (95% CI), nsP > 0.05, ****P < 0.0001. Shown are mean values ± SEM of biological triplicates. i VSV∆51-amiR-NTC or VSV∆51-amiR-4 titers of ex vivo infected tumour cores obtained from several pancreatic cancer PDXs. Results displayed as mean ± SEM (P014, n = 10; P020, n = 40; P021, n = 29; P025, n = 52; P027, n = 10), two-tailed unpaired t test of individual patient plots, nsP > 0.05, *P = 0.0213, **P = 0.007. j Immunohistochemistry staining of ARID1A protein expression in five pancreatic cancer patient-derived xenografts (PDXs). Scale bar = 100 μm; (n = 2 technical replicates per PDX). k Gene-Ontology (GO) analysis of biological processes differentially expressed between PANC1 wild-type and ARID1A-knockout cells. Illustrated GO-terms represent all significantly different biological processes (Fisher’s exact test) after correction for multiple hypothesis testing (FDR). l Heatmap showing gene transcript expression levels (Log2 TPM [transcripts per kilobase million]) of antiviral genes in uninfected PANC1 wild-type or ARID1A-knockout cells. Source data are provided as a Source data file.

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