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. 2023 Jul 22;4(4):e337. doi: 10.1002/mco2.337

FIGURE 4.

FIGURE 4

Knockout of ubiquitin‐specific protease 5 (USP5) promotes ferroptosis. (A) Representative phase‐contrast images of Hep3B cells knockout of USP5 treated with RSL‐3 (10 μM, 24 h). (B) Analysis of the responses of Hep3B cells knockout of USP5 to Fer‐1 (10 μM, 24 h) and RSL‐3 (10 μM, 24 h) was done using cell counting kit‐8 (CCK8) assays. (C) The expression level of lymphoid‐specific helicase (LSH) and ferroptosis‐related proteins were detected using western blot in Hep3B cells knockout of USP5. (D and E) The levels of total GSH (D) and lipid ROS (E) in Hep3B cells knockout of USP5 were analyzed. (F) Western blot was used to detect the expression level of LSH and SLC7A11 in Hep3B cells knockout of USP5 with LSH overexpression. (G) Representative phase‐contrast images of Hep3B cells knockout of USP5 with LSH overexpression treated with RSL‐3 (10 μM, 24 h). (H and I) The levels of total GSH (H) and lipid ROS (I) in Hep3B cells knockout of USP5 with LSH overexpression were analyzed. Ns, nonsignificant (p > 0.05), *p < 0.05, **p < 0.01, and ***p < 0.001.