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. 2020 Mar 4;217(5):e20191174. doi: 10.1084/jem.20191174

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© 2020 Cai et al.

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Figure 6. — USP22-deficient mice exhibit increased susceptibility to viral infection. (A and F) Cre-ER Usp22^fl/+ (n = 14) and Cre-ER Usp22^fl/fl (n = 14) mice were intraperitoneally injected with tamoxifen (80 µg/g dissolved in corn oil) for 5 consecutive days and rested for 7 d followed by various experiments. Survival (Kaplan–Meier curve) analysis of these mice intravenously injected with VSV (2 × 10⁷ PFU per mouse; A) or HSV-1 (3 × 10⁶ PFU per mouse; F). (B and G) ELISA analysis of IFN-β and CCL5 in the sera of Cre-ER Usp22^fl/+ and Cre-ER Usp22^fl/fl mice treated as in A and intravenously injected with VSV (2 × 10⁷ PFU per mouse, n = 6; B) or HSV-1 (3 × 10⁶ PFU per mouse, n = 6; G) for 12 h. (C, D, H, and I) qRT-PCR analysis of Ifnb, Isg15, Ccl5, or Usp22 mRNA in the lungs (C and H, n = 6 per group) and brains (D and I, n = 6 per group) from Cre-ER Usp22^fl/+ and Cre-ER Usp22^fl/fl mice treated as in A followed by intravenous injection with VSV (2 × 10⁷ PFU per mouse) or HSV-1 (5 × 10⁶ PFU per mouse) for 24 h (C and H) or 4 d (D and I). (E and J) Plaque assays analyzing VSV or HSV-1 titers in the brains and kidneys (E) or brains and spleens (J) from Cre-ER Usp22^fl/+and Cre-ER Usp22^fl/fl mice treated as in A followed by intraperitoneal injection with VSV (2 × 10⁷ PFU per mouse, n = 6) or HSV-1 (5 × 10⁶ PFU per mouse, n = 6) for 4 d. (K) Survival (Kaplan–Meier curve) analysis of Lyz2-Cre Usp22^fl/+ (n = 9, left; n = 10, right) and Lyz2-Cre Usp22^fl/fl (n = 10, left; n = 9, right) mice intravenously injected with VSV (2 × 10⁷ PFU per mouse; left) or HSV-1 (3 × 10⁶ PFU per mouse; right). (L) ELISA analysis of IFN-β and CCL5 in the sera of Lyz2-Cre Usp22^fl/+ and Lyz2-Cre Usp22^fl/fl mice intravenously injected with VSV (2 × 10⁷ PFU per mouse, n = 5) or HSV-1 (3 × 10⁶ PFU per mouse, n = 5) for 12 h. *, P < 0.05; **, P < 0.01; and ***, P < 0.001 (Student’s t test). Data are a combination (A, F, and K) or representative (B–E, G–J, L, and M) of two independent experiments (graphs show mean ± SD).