Fig 1: Loss of ING3 induces robust IFN and ISG expression. (A) Volcano plots of RNA-sequencing analysis of WT and ING3 KO (clones #1 and #2) HT-29 cells. Genes that were up-regulated in ING3 KO cells are highlighted in red, and the highly induced ISGs were marked with more prominent red dots (Padj: adjusted p-value). (B) IFI44L and IFITM1 transcript levels were measured by RT-qPCR and normalized to that of GAPDH in WT and ING3 KO HT-29 cells. (C) IFNL3 transcript level was measured by RT-qPCR and normalized to that of GAPDH in WT and ING3 KO HT-29 cells transduced with lentiviruses in empty vector (ev), encoding mGFP-ING3, or encoding Myc-ING3. (D) Same as (C) except that cell culture supernatants were collected and secreted IFN-λ levels were measured by ELISA. (E) WT and ING3 KO HT-29 cells were treated with vehicle control DMSO or JAK inhibitor ruxolitinib (100 nM) for 24 hr and infected with RV (MOI = 1) for another 24 h. RV NSP5 level was measured by RT-qPCR and normalized to that of GAPDH. For all panels except A, experiments were repeated at least three times with similar results. Figure A was performed once with technical duplicates. Raw data is listed in Dataset S1. Data are represented as mean ± SEM. Statistical significance is from pooled data of the multiple independent experiments (**P ≤ 0.01; ***P ≤ 0.001).