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. 2024 Oct 1;15:8479. doi: 10.1038/s41467-024-52773-w

Fig. 7. Loss or inhibition of TRPM7 is protective against SARS-CoV-2 and other coronavirus infection.

Fig. 7

a Schematic of VSV-coronavirus chimera construct with genetically encoded eGFP that is transcribed upon infection. Different variant-specific spike proteins were expressed as the envelope glycoprotein. b Maximum projection of cells infected with VSV-SARS-CoV-2-eGFP variants (green). Vero cells +/- TMPRSS2 were transfected with Control (Ctl) and TRPM7 siRNA (M7KD). Cells were then infected with VSV-SARS-CoV-2-eGFP expressing the Spike protein of the Wuhan, Delta, or Omicron variants. Cells were incubated with virus for 8 hours before fixation and imaging. The scale bar at the bottom left of the images is representative of 50 µm for all images across each row. c Quantification VSV-SARS-CoV-2-eGFP variant infections in control (gray) and M7KD (blue) Vero cells. Cells were infected with VSV pseudotyped with Wuhan, Delta, and Omicron spike variants. P values determined by two-tailed Student’s t test. n = 3 independent experiments. Mean and SEM plotted. d Quantification of infection in Vero cells stably overexpressing TMPRSS2 with VSV-SARS-CoV-2-eGFP variants in control (gray) and M7KD (blue) cells. P values determined by two-tailed Student’s t test. n = 3 independent experiments. Mean and SEM is plotted. e Maximum projection (20 μm) of Vero and Vero-TMPRSS2 cells infected with VSV-SARS-CoV-2-eGFP Wuhan strain in the presence of 5 μM FTY720 or 30 μM NS8593. Cells were pretreated with FTY720 and NS8593 for 15 minutes before exposure to virus and throughout the infection assay. f Quantification of VSV-SARS-CoV-2-eGFP infection in Vero and Vero-TMPRSS2 cells in the presence of 5 μM FTY720 (purple) or 30 μM NS8593 (teal) or without drug (gray). P values were determined by one-way ANOVA with Tukey’s multiple comparison test. n = 3 independent experiments. Mean and SEM plotted. g Quantification of infection by VSV-MERS, VSV-SARS-CoV-1, and VSV-SARS-CoV-2 Wuhan variant in Control (Ctl), TRPM7 (M7KD), TPC1 (TPC1KD), and TPC2 (TPC2KD) knockdown Vero cells. Infection assays were performed in parental Vero cells and Vero cells stably overexpressing TMPRSS2. P values determined by one-way ANOVA with Tukey’s multiple comparison test. n = 3 independent experiments. Mean and SEM plotted.