Figure 1. FEZ1 regulates ISG expression in microglia cells.

(A and B) siRNA-mediated depletion of FEZ1 increases expression of ISGs (MxA, MxB, PKR, and ISG56) in CHME3s infected with HSV-1 (A) or VacV (B) as detected by the expression of viral proteins (HSV1: ICP4, ICP, ICP5; VacV: I3, G8, A25). L.E., long exposure; S.E., short exposure.

(C) KO of FEZ1 (FEZ1 1–4 or FEZ1 pooled), but not non-targeting (NT) gRNAs, increases ISG levels in CHME3s.

(D) Quantification of the ISG levels relative to HSPA8 in FEZ1 KO CHME3s from (C). Data are presented as the ratio to the difference between control and treatment groups.

(E) WB analysis showing effects of FEZ1-Flag or S58A-Flag on expression of ISGs in CHME3s.

(F) Quantification of ISG levels relative to HSPA8 in control, FEZ1-Flag, or FEZ1-S58A-Flag expressing CHME3s from (E). Statistical significance is presented as “A” when it was calculated for all groups using one-way ANOVA followed by Dunnett post-hoc test or as “t” if Student’s t test was applied to compare pairwisely Flag or FEZ1-S58A-Flag with FEZ1-Flag.

(G and H) Measurement of IFN-β levels in culture medium of CHME3 expressing Flag, FEZ1-Flag, and S58A-Flag by ELISA (G). Culture medium of CHME3s spiked with IFN-β was included as positive control. Note that samples (except for the spiked control) were concentrated in order to obtain readings within the range of the standard curve (H) and are divided accordingly to present the actual values shown. One-way ANOVA followed by Tukey post-hoc test was used to calculate statistical significance in (D and G). (D, F and G) n = 3; red line, mean; bars, SD.

See also Figure S1.