HCMV infection downregulates GPX4 and stimulates lipid peroxidation but does not induce ferroptosis Figure 4

GPX4 knockdown inhibits viral infection and increases ferroptosis markers. (A) Western blot analysis of MRC-5 cells expressing control shRNA (shGFP) or GPX4 shRNA (shGPX4-1 and shGPX4-2). Cells were either uninfected or infected with TB40/E-WT at MOI = 3 and harvested at 24 or 96 hpi. Blots were probed for GPX4, IE2, and p115 as a loading control. The values under the GPX4 blot denote quantification of each GPX4 band in the blot, relative to the GPX4 band in shGFP uninfected cells. (B) Infectious titers at 120 hpi from HCMV-infected (as described in panel A) MRC-5 cells expressing shRNA against green fluorescent protein (GFP) or GPX4. Results are from five independent experiments, n = 15. (C) Lipid peroxidation measured by C11 BODIPY FITC fluorescence shift in MRC-5 cells expressing shGFP or either shGPX4 clone either uninfected or infected with TB40/E-WT (MOI = 3) at 96 hpi. Data are shown relative to control (shGFP) uninfected cells. Results are from two independent experiments, n = 6. (D) ATP levels in MRC-5 cells expressing control shGFP or shGPX4, either uninfected or HCMV-infected (as described in panel A) and measured at 96 hpi. ATP levels are shown relative to respective uninfected or infected shGFP-expressing cells. Results are from two independent experiments n = 8. (E) LDH release in uninfected MRC-5 cells expressing shRNA against GFP or GPX4. LDH release is shown as the percent of maximum LDH in uninfected control cells. Results are from two independent experiments, n = 6. (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. Results are from two independent experiments, n = 6. For the above panels, * indicates P < 0.05 and ns = not significant.

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Norbury, Christopher (2024). HCMV infection downregulates GPX4 and stimulates lipid peroxidation but does not induce ferroptosis Figure 4 [Data set]. Scholarsphere.

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Work Title HCMV infection downregulates GPX4 and stimulates lipid peroxidation but does not induce ferroptosis Figure 4
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  1. Christopher Norbury
License CC BY 4.0 (Attribution)
Work Type Dataset
Publication Date December 20, 2024
Deposited December 20, 2024

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  • Updated Description, Publication Date Show Changes
    Description
    • GPX4 knockdown inhibits viral infection and increases ferroptosis markers. (A) Western blot analysis of MRC-5 cells expressing control shRNA (shGFP) or GPX4 shRNA (shGPX4-1 and shGPX4-2). Cells were either uninfected or infected with TB40/E-WT at MOI = 3 and harvested at 24 or 96 hpi. Blots were probed for GPX4, IE2, and p115 as a loading control. The values under the GPX4 blot denote quantification of each GPX4 band in the blot, relative to the GPX4 band in shGFP uninfected cells. (B) Infectious titers at 120 hpi from HCMV-infected (as described in panel A) MRC-5 cells expressing shRNA against green fluorescent protein (GFP) or GPX4. Results are from five independent experiments, n = 15. (C) Lipid peroxidation measured by C11 BODIPY FITC fluorescence shift in MRC-5 cells expressing shGFP or either shGPX4 clone either uninfected or infected with TB40/E-WT (MOI = 3) at 96 hpi. Data are shown relative to control (shGFP) uninfected cells. Results are from two independent experiments, n = 6. (D) ATP levels in MRC-5 cells expressing control shGFP or shGPX4, either uninfected or HCMV-infected (as described in panel A) and measured at 96 hpi. ATP levels are shown relative to respective uninfected or infected shGFP-expressing cells. Results are from two independent experiments n = 8. (E) LDH release in uninfected MRC-5 cells expressing shRNA against GFP or GPX4. LDH release is shown as the percent of maximum LDH in uninfected control cells. Results are from two independent experiments, n = 6. (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. Results are from two independent experiments, n = 6. For the above panels, * indicates P < 0.05 and ns = not significant.
    Publication Date
    • 2024-12-20
  • Added Creator Christopher Norbury
  • Added Maddie Martin 2023-10-13 14h57m12s.scn
  • Added Maddie Martin 2023-10-13 11h27m08s.scn
  • Added 082323 GPX4 KD titers.xlsx
  • Added 063023 GPX4 FSP1 KD titers .xlsx
  • Added 090123 FSP1 & GPX4 KD titers.xlsx
  • Added 092823 GPX4 KD titer and TB40WT retiter.xlsx
  • Added 101323 GPX4 KD C11 BODIPY combined.xlsx
  • Added 101323 GPX4 KD JC-10 combined.xlsx
  • Added 101323 GPX4 KD LDH Combined.xlsx
  • Updated
  • Added Figure 4.pdf
  • Added Fig4 readme.txt
  • Updated License Show Changes
    License
    • https://creativecommons.org/licenses/by/4.0/
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