
HCMV infection downregulates GPX4 and stimulates lipid peroxidation but does not induce ferroptosis Figure 5
FSP1 knockdown does not impair infection or induce ferroptosis. (A) Western blot analysis of lysates from uninfected MRC-5 cells expressing control shRNA (green fluorescent protein [GFP]) or shFSP1 (shRNA against FSP1). Blots were probed for FSP1 and p115 as a loading control. The values under the blot denote quantification of each FSP1 band in the blot, relative to the FSP1 band in shGFP cells. (B) Infectious titers at 120 hpi from MRC-5 cells expressing GFP control shRNA or shFSP1 and infected with TB40/E-WT at MOI = 3. Results are from seven independent experiments, n = 18. (C) Lipid peroxidation was measured by C11 BODIPY FITC fluorescence shift in MRC-5 cells expressing shGFP or shFSP1 and either uninfected or infected as described in panel B at 96 hpi. MRC-5 cells treated with RSL3 (1 μM) were used as a positive control. Data are shown relative to uninfected shGFP cells. Results are from two independent experiments, n = 6. (D) ATP levels in MRC-5 cells infected as described in panel B at 96 hpi with cells expressing scrambled shRNA or shFSP1. Data are shown relative to shSCR cells. Results are from two independent experiments, n = 9. (E) LDH release in uninfected MRC-5 cells expressing shRNA against GFP or FSP. LDH release is shown as the percent of maximum LDH in uninfected control cells. Results are from three independent experiments, n = 9. (F) LDH release in MRC-5 cells expressing shRNA against GFP or GPX4 and infected with HCMV (as in panel A) and measured at 96 hpi. LDH release is shown as the percent of maximum LDH in infected cells. For the above panels, * indicates P < 0.05 and ns = not significant. Results are from three independent experiments, n = 9. For the above panels, * indicates P < 0.05 and ns = not significant.
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Work Title | HCMV infection downregulates GPX4 and stimulates lipid peroxidation but does not induce ferroptosis Figure 5 |
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License | CC BY 4.0 (Attribution) |
Work Type | Dataset |
Publication Date | December 20, 2024 |
Deposited | December 20, 2024 |