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. 2022 May 24;14(11):2580. doi: 10.3390/cancers14112580

Figure 2.

Figure 2

MUC1-C is necessary for induction of RNA PRRs. (A–D) RNA-seq was performed in triplicate on BT-549 cells expressing a tet-MUC1shRNA, which were treated with vehicle or DOX for 7 days. Candidate pathway enrichment plots for the HALLMARK INTERFERON ALPHA RESPONSE (A), HALLMARK INFLAMMATORY RESPONSE (B), KEGG RIG-I LIKE RECEPTOR SIGNALING (C) and REACTOME DDX58 IFIH1 MEDIATED INDUCTION OF INTERFERON ALPHA BETA (D) gene signatures. (E,F) BT-549/tet-MUC1shRNA cells were treated with vehicle or DOX for 7 days. MUC1-C, RIG-I and MDA5 mRNA levels were analyzed by qRT-PCR (E). The results (mean ± SD of 4 determinations) are expressed as relative mRNA levels compared to that obtained for vehicle-treated cells (assigned a value of 1). The asterisk (*) denotes a p-value < 0.05. Lysates were immunoblotted with antibodies against the indicated proteins (F, left). Lysates from BT-549/CshRNA and BT-549/MUC1shRNA#2 were immunoblotted with antibodies against the indicated proteins (F, right). (G,H). BT-549/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days and stimulated with poly I:C for 12 h (G) or IFN-β for 4 h (H) were analyzed for RIG-I and MDA5 mRNA levels by qRT-PCR. The results (mean ± SD of 4 determinations) are expressed as relative mRNA levels compared to that obtained for vehicle-treated cells (assigned a value of 1). Uncropped Western Blots can be found at supplemental original images.