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.

Citation

Norbury, Christopher (2024). HCMV infection downregulates GPX4 and stimulates lipid peroxidation but does not induce ferroptosis Figure 3 [Data set]. Scholarsphere.

Files

Metadata

Work Title HCMV infection downregulates GPX4 and stimulates lipid peroxidation but does not induce ferroptosis Figure 3
Access
Open Access
Creators
  1. Christopher Norbury
License CC BY 4.0 (Attribution)
Work Type Dataset
Publication Date December 20, 2024
Deposited December 20, 2024

Versions

Analytics

Collections

Work History

Version 1
published

  • Created
  • Updated
  • Updated Description, Publication Date Show Changes
    Description
    • 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.
    Publication Date
    • 2024-12-20
  • Added Creator Christopher Norbury
  • Added combined RSL3, Erastin titers.xlsx
  • Added RSL3, Erastin titers combined.xlsx
  • Added 022323 RSL3, Erastin titers redo.xlsx
  • Added 021523 iFSP1, RSL3, Erastin titer plates H15 and D4.xlsx
  • Added 090823 GSH Assay.xlsx
  • Added 092623 GSH Assay.xlsx
  • Added 100423 GSH Assay.xlsx
  • Added 110823 GSH assay.xlsx
  • Added 111423 GSH assay.xlsx
  • Added 111723 GSH assay.xlsx
  • Added 012423 Cyquant LDH Assay iFSP1, RSL3, Erastin.xlsx
  • Added 022823 Cyquant LDH Assay iFSP1, RSL3, Erastin.xlsx
  • Added 031023 Cyquant LDH Assay RSL3, Erastin treatment.xlsx
  • Added 050223 RSL3 erastin cell titer glo.xlsx
  • Added 020323 Ferr Inducers CellTiterGlo.xlsx
  • Added 031023 cell titer glo RSL3 4h, Erastin 14h treatment.xlsx
  • Updated License Show Changes
    License
    • https://creativecommons.org/licenses/by/4.0/
  • Added Figure 3.pdf
  • Added Fig3 readme.txt
  • Published
  • Updated