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. Author manuscript; available in PMC: 2021 Apr 8.
Published in final edited form as: Cell Host Microbe. 2020 Mar 5;27(4):556–570.e6. doi: 10.1016/j.chom.2020.02.004

Figure 5. TMEM173-Mediated GSDMD Cleavage Promotes F3 Release.

Figure 5.

(A–C) Western blot analysis of GSDMD-N production in indicated knockout (A), knockdown (B), or mutated (C) THP1 cells after E. coli (25 MOI) or S. pneumoniae (25 MOI) infection for 24 h.

(D and E) Analysis of GSDMD-N production (D) and F3 release (E) in indicated BMDM cells after E. coli (25 MOI) or S. pneumoniae (25 MOI) infection for 24 h in the absence or presence of the CASP8 inhibitor Z-IETD-FMK (20 μM) (n = 3 wells per group; *p < 0.05 versus control group, t test).

(F) Cell lysates were analyzed by anti-FLAG antibody in Tmem173−/− THP1 cells after transfection indicated GSDMD mutants.

(G) Analysis of F3 release in indicated THP1 cells after E. coli (25 MOI) or S. pneumoniae (25 MOI) infection for 24 h in the absence or presence of the PLC inhibitor U73122 (10 μM) (n = 3 wells per group; *p < 0.05, t test).

(H) Western blot analysis of protein expression in THP1 cells after E. coli (25 MOI) or S. pneumoniae (25 MOI) infection for 24 h.

(I) Western blot analysis of protein expression in indicated THP1 cells after E. coli (25 MOI) or S. pneumoniae (25 MOI) infection in the absence or presence of BAPTA-AM (10 μM) for 24 h.

Data are presented as mean ± SD ([E] and [G]).