
HCMV infection downregulates GPX4 and stimulates lipid peroxidation but does not induce ferroptosis Figure 3
FIG 3 Reduced levels of GPX4 impair infection and induce ferroptosis. (A) Infectious titers at 120 hpi from TB40/E-WT (MOI = 3) infected MRC-5 cells treated with vehicle control (DMSO), erastin (5 μM), or RSL3 (1 μM) at 72 hpi. Results are from three independent experiments, n = 4. (B) Lipid peroxidation was measured with C11 BODIPY FITC fluorescence shift in MRC-5 cells either uninfected or infected (as in [A]), treated with DMSO, erastin, or RSL3, and harvested at 96 hpi. Infected cells were treated for 24 hours before harvest, while uninfected cells were treated for 14 hours (DMSO and erastin) or 4 hours (RSL3). Lipid peroxidation is shown relative to uninfected, DMSO-treated cells. Results are from three independent experiments, n = 6. (C) ATP levels at 96 hpi in uninfected MRC-5 cells treated with DMSO, erastin, or RSL3 as described in (A), or MRC-5 cells infected with TB40/E-WT and treated as described in (A). Data are shown as %ATP relative to DMSO-treated cells in the uninfected or HCMV-infected group. Results are from two independent experiments, n = 6. (D) LDH release in uninfected MRC-5 cells treated with DMSO, erastin, or RSL3 for 24 hours. LDH release is shown as the percent of maximum LDH in uninfected cells. Results are from three independent experiments, n = 9. (E) LDH release in MRC-5 cells infected with HCMV (as in [A]) and treated with DMSO, erastin, or RSL3 at 72 hpi and measured at 96 hpi. LDH release is shown as the percent of total LDH in infected cells. Results are from three independent experiments, n = 9. (F) GSH levels measured in uninfected or TB40/E-WT (MOI = 3) infected MRC-5 cells at 96 hpi following treatment with DMSO, erastin, or RSL3. Infected cells were treated for 24 hours before harvest, while uninfected cells were treated for 14 hours (DMSO and erastin) or 4 hours (RSL3). Results are from three independent experiments, n = 8. 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 3 |
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License | CC BY 4.0 (Attribution) |
Work Type | Dataset |
Publication Date | December 20, 2024 |
Deposited | December 20, 2024 |