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. 2020 Sep 14;39(22):e104106. doi: 10.15252/embj.2019104106

Figure 1. STING‐mediated induction of a subset of genes needs EGFR kinase activity.

Figure 1

  • A
    Raw 264.7 cells were pretreated with DMSO or the EGFR inhibitor gefitinib (10 μM) before cGAMP transfection; after 5 h, IFN‐β mRNA was measured by qRT–PCR (n = 3). Bars are the mean ± SD of indicated (n) independent experiments. ***P < 0.001 when compared to gefitinib‐pretreated group by two‐tailed Student's t‐test.
  • B
    WT and EGFR knockdown Raw 264.7 cells were transfected with cGAMP; 4.5 h post‐treatment, IFN‐β mRNA was measured (n = 3). Bars are the mean ± SD of indicated (n) independent experiments. ***P < 0.001 when compared to gefitinib‐treated group by two‐tailed Student's t‐test.
  • C
    WT and EGFR−/− BMDMs were transfected with cGAMP; 5 h post‐transfection, IFN‐α mRNA induction was measured (n = 4). Bars are the mean ± SD of indicated (n) independent experiments. ***P < 0.001 when compared to EGFR−/− cells by two‐tailed Student's t‐test.
  • D
    WT and EGFR−/− HeLa cells were transfected with cGAMP; after 5 h, IFN‐β mRNA was measured (n = 3).
  • E–G
    MEF cells were pretreated with gefitinib for 1 h and then transfected with cGAMP; 4.5 h post‐treatment, total RNA was harvested for analysis. (E) RNA‐seq analysis (n = 2). For RNA‐seq analysis, poly‐T-enriched cDNA library was prepared and 30 M single‐end 50‐bp reads were analyzed. The heat map for some of the genes is shown. (F) 259 mRNAs were induced by twofold or more upon cGAMP treatment; the induction of 86 of those genes was inhibited by gefitinib by twofold or more. (G) The RNA‐seq analysis results were confirmed by qRT–PCR analysis of a gefitinib‐sensitive and an gefitinib‐insensitive mRNA (n = 3). Bars are the mean ± SD of indicated (n) independent experiments. ***P < 0.001 when compared to gefitinib‐treated group by two‐tailed Student's t‐test.
  • H
    MEF cells were pretreated with gefitinib for 1 h and followed with cGAMP or Poly (I:C) transfection; 4.5 h post‐treatment, total RNA was harvested and IFN‐α mRNA induction was measured (n = 3). Bars are the mean ± SD of indicated (n) independent experiments. ***P < 0.001 when compared to gefitinib‐pretreated group by two‐tailed Student's t‐test; NS means no significance.