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. Author manuscript; available in PMC: 2022 Mar 7.
Published in final edited form as: Cell Rep. 2022 Feb 15;38(7):110396. doi: 10.1016/j.celrep.2022.110396

Figure 4. FEZ1 and HSPA8 regulate nuclear localization of DNA-PK.

Figure 4.

(A, C, and E) Representative IF images of DNA-PK staining in three different FEZ1 KO CHME3 pools (FEZ1 KO 1, 3, and 4) (A), or CHME3s expressing Flag control, or FEZ1-Flag or S58A-Flag (C), or HSPA8 KO CHME3 pools (HSPA8 KO 1, 2, and 4) (E). Nuclei were stained with Hoechst (blue). Scale bar, 10 μm.

(B, D, and F) Quantitative analysis of nuclear DNA-PK staining in samples from (A, C, and E), respectively.

(G–L) Responses to CT-DNA in CHME3 microglia. (G) WB analysis showing decreased phosphorylated FEZ1 at S58 (pFEZ1) as well as increased levels of phosphorylated IRF3 (pIRF3) and ISGs (MxA, MxB, and ISG56) in CHME3s treated with CT-DNA. (H) Quantification of the pFEZ1 levels and ISGs from samples in (G) n = 3; bars, SD. (I–L) Representative images (I and K) and quantitative analysis (J and L) of HSPA8 (I and J) or DNA-PK (K and L) staining in nuclei of CHMEs either untreated or treated with CT-DNA. One-way ANOVA followed by Dunnett post-hoc test (B and F) or Tukey post-hoc test (D), and t test (J, H, and L) was used to calculate statistical significance. (B, D, F, J, and L) Number of cells analyzed is indicated; n = 3; red line, mean; bars, SD.

See also Figures S4 and S5.