Figure 5. Knockdown of TRIM65 suppresses MDA5 activation in human cells. (A) qPCR analysis of TRIM65 expression in HeLa cells stably expressing shRNA against TRIM65. (B and C) qPCR analysis of IFN-β expression in HeLa cells stably expressing shRNA against TRIM65 and left stimulated for 6 h with cytosolic HMW poly I:C (B), LMW poly I:C (B), EMCV-RNA (C), or VSV-RNA (C). (D) Immunoblot analysis of TRIM65 expression in HeLa cells stably expressing shRNA against TRIM65 with or without overexpression of GFP-TRIM65. (E) qPCR analysis of IFN-β expression in HeLa cells stably expressing shRNA against TRIM65 with or without overexpression of GFP-TRIM65 and left stimulated for 6 h with cytosolic EMCV-RNA. Data are from three independent experiments with biological duplicates in each (mean and SEM of n = 6). Statistics were analyzed via an unpaired Student’s t test: ***, P < 0.001.
Figure 5.

Knockdown of TRIM65 suppresses MDA5 activation in human cells. (A) qPCR analysis of TRIM65 expression in HeLa cells stably expressing shRNA against TRIM65. (B and C) qPCR analysis of IFN-β expression in HeLa cells stably expressing shRNA against TRIM65 and left stimulated for 6 h with cytosolic HMW poly I:C (B), LMW poly I:C (B), EMCV-RNA (C), or VSV-RNA (C). (D) Immunoblot analysis of TRIM65 expression in HeLa cells stably expressing shRNA against TRIM65 with or without overexpression of GFP-TRIM65. (E) qPCR analysis of IFN-β expression in HeLa cells stably expressing shRNA against TRIM65 with or without overexpression of GFP-TRIM65 and left stimulated for 6 h with cytosolic EMCV-RNA. Data are from three independent experiments with biological duplicates in each (mean and SEM of n = 6). Statistics were analyzed via an unpaired Student’s t test: ***, P < 0.001.

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