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. Author manuscript; available in PMC: 2022 Jan 1.
Published in final edited form as: Cancer Res. 2021 May 3;81(13):3607–3620. doi: 10.1158/0008-5472.CAN-20-3790

Figure 3. dsRNA sensing MDA5-NFκB signaling axis leads to type I IFN production in DDX3X-depleted breast cancer cells.

Figure 3.

A. Western blot analysis of STING, phosphorylation of STING, and cGAS in DDX3X-control or -KD MCF7 or MDA-MB-453 cells.

B. Western blot analysis of phosphorylation of TBK1, IRF3, and IRF7 in DDX3X-control or -KD MCF7 or MDA-MB-453 cells.

C. Western blot analysis of phosphorylation of NFκB p65 and IκBα in DDX3X-control or -KD MCF7 or MDA-MB-453 cells.

D. qRT-PCR of RIG-I, MDA5, and OAS1 in DDX3X-control or KD-MCF7 cells.

E. Western blot analysis of RIG-I, MDA5, and OAS1 in DDX3X-control or -KD MCF7 or MDA-MB-453 cells (left). Bar graphs of relative protein band intensity (right).

F. Western blot validation of CRISPR/Cas9-mediated MDA5 knockout (KO) in DDX3X-control or -KD MCF7 cells (left). qRT-PCR of IFNB1 in MDA5 wildtype or KO cells followed by DDX3X-control or -KD.

G. Colony formation assay in CRISPR/Cas9-mediated MDA5-KO MCF7 cells followed by DDX3X-control or -KD.

Data are shown as mean ± SEM of three independent experiments. Statistical significance was calculated using unpaired t-tests (Fig. 3D and 3E) or one-way ANOVA with uncorrected Fisher’s LSD test (Fig. 3F). *p < 0.05; **p< 0.01.