Fig. 4.

Knockdown of CALCOCO2 results in increased protein levels of MAVS and p-TBK1 as well as elevated production of type I interferons. a, c, e HeLa cells were treated with siCALCOCO2 or siCON for 48 h. Cell lysates were harvested and probed for MAVS (a) and p-TBK1/TBK1 (c) by western blotting. Knockdown efficiency was verified by probing with anti-CALCOCO2 antibody. Densitometry was carried out as in Fig. 1. Total RNA was extracted and real-time qRT-PCR was performed to measure mRNA levels of IFN-α and IFN-β normalized to GAPDH mRNA (e), and presented as relative quantification (RQ) with respect to siCON (RQ ± SE, n = 3, confidence interval at 95%). b, d, f HeLa cells were transfected with siSQSTM1 or siCON for 48 h, followed by western blot analysis of MAVS (b) and p-TBK1/TBK1 (d) and qRT-PCR measurement of IFN-α and IFN-β (f) as above. Similar results were observed in three independent experiments