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The viability of primary CD4 + T cells was assessed by the MTT assay following treatment with DMSO or the indicated concentration of METTL3 inhibitors for ( A ) 24 h and ( B ) 48 h. Results are shown as mean ± SD from three independent donor experiments. Statistical analysis was performed using two-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001 and **** P < 0.0001. Statistical significance was compared between treated samples and the DMSO control.

Journal: bioRxiv

Article Title: Pharmacological METTL3 inhibition attenuates HIV-1 latency reversal in CD4 + T cells

doi: 10.64898/2026.03.18.712554

Figure Lengend Snippet: The viability of primary CD4 + T cells was assessed by the MTT assay following treatment with DMSO or the indicated concentration of METTL3 inhibitors for ( A ) 24 h and ( B ) 48 h. Results are shown as mean ± SD from three independent donor experiments. Statistical analysis was performed using two-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001 and **** P < 0.0001. Statistical significance was compared between treated samples and the DMSO control.

Article Snippet: J-Lat 10.6 and primary CD4 + T cells were cultured in RPMI-1640 (ATCC) supplemented with 10% fetal bovine serum (FBS; R&D systems) and antibiotics (100 U/mL penicillin and 100 μg/mL streptomycin, Gibco).

Techniques: MTT Assay, Concentration Assay, Control

( A ) Protocol summary. Naïve CD4 + T cells were isolated from PBMCs of three healthy donors. Cells were activated using anti-CD3/CD28 antibody-coated beads for 3 days and infected with single-cycle HIV-1-GFP reporter at day 7 to generate a primary T CM model of latency. ( B ) GFP expression was measured at day 10 and 22 by flow cytometry. Representative data from a single donor are shown. SSC, side scatter. The percentage of GFP-positive cells are shown on the plots. ( C ) After 10 days of HIV-1 infection and culture, cells were treated with either DMSO or METTL3 inhibitors and m 6 A levels of cellular RNA were measured by ELISA. The relative m 6 A levels were normalized to the DMSO control that was set as 1. Results from three individual donors are shown. ( D and E ) HIV-1 latency reversal was measured by GFP expression using flow cytometry at day 22. Changes in (D) GFP percentage and (E) mean fluorescence intensity (MFI) of GFP-positive cells from three independent donors are shown. Statistical analysis was performed using two-way ANOVA. * P < 0.05, and *** P < 0.001.

Journal: bioRxiv

Article Title: Pharmacological METTL3 inhibition attenuates HIV-1 latency reversal in CD4 + T cells

doi: 10.64898/2026.03.18.712554

Figure Lengend Snippet: ( A ) Protocol summary. Naïve CD4 + T cells were isolated from PBMCs of three healthy donors. Cells were activated using anti-CD3/CD28 antibody-coated beads for 3 days and infected with single-cycle HIV-1-GFP reporter at day 7 to generate a primary T CM model of latency. ( B ) GFP expression was measured at day 10 and 22 by flow cytometry. Representative data from a single donor are shown. SSC, side scatter. The percentage of GFP-positive cells are shown on the plots. ( C ) After 10 days of HIV-1 infection and culture, cells were treated with either DMSO or METTL3 inhibitors and m 6 A levels of cellular RNA were measured by ELISA. The relative m 6 A levels were normalized to the DMSO control that was set as 1. Results from three individual donors are shown. ( D and E ) HIV-1 latency reversal was measured by GFP expression using flow cytometry at day 22. Changes in (D) GFP percentage and (E) mean fluorescence intensity (MFI) of GFP-positive cells from three independent donors are shown. Statistical analysis was performed using two-way ANOVA. * P < 0.05, and *** P < 0.001.

Article Snippet: J-Lat 10.6 and primary CD4 + T cells were cultured in RPMI-1640 (ATCC) supplemented with 10% fetal bovine serum (FBS; R&D systems) and antibiotics (100 U/mL penicillin and 100 μg/mL streptomycin, Gibco).

Techniques: Isolation, Infection, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Control, Fluorescence