Fig. 5: TOLLIP is involved in ACE2 degradation and the associated SARS-CoV-2 infection. | Nature Communications

Fig. 5: TOLLIP is involved in ACE2 degradation and the associated SARS-CoV-2 infection.

From: Suppression of ACE2 SUMOylation protects against SARS-CoV-2 infection through TOLLIP-mediated selective autophagy

Fig. 5

a Immunoprecipitation and immunoblot analysis of 293 T cells transfected with vectors of HA-ACE2 and indicated Flag-tagged cargo receptors. b Representative ACE2 (red) and TOLLIP (green) immunofluorescent staining in lung from transgenic hACE2 mice upon SARS-CoV-2 (1 × 105 TCID50/mL) infection for 72 h from n = 5 independent biological mice per group. Scale bar, 50 μm. c Area of ACE2 and TOLLIP colocalization. d Calu-3 cells were challenged with SARS-CoV-2 (MOI = 0.5) for indicated time points and the protein extracts were immunoprecipitated and analyzed by immunoblotting. e Calu-3 cells transfected with scramble or TOLLIP-specific siRNAs were infected with SARS-CoV-2 S pseudotyped virus for 48 h, the protein extracts were harvested for luciferase assay. f Immunoblot analysis of Calu-3 cells transfected with scramble or TOLLIP-specific siRNAs and infected with SARS-CoV-2 (MOI = 0.5) for indicated time points. g Plaque titration of SARS-CoV-2 in supernatants of scrambled or TOLLIP-specific siRNAs transfected Calu-3 cells infected with SARS-CoV-2 (MOI = 0.5). h GSEA analysis of genes expressed differentially in Calu-3 cells with or without TOLLIP-specific siRNA transfection upon SARS-CoV-2 (MOI = 0.5) infection for 24 h. FDR (q-value) was shown. i Heatmap indicating the transcription levels of DEGs induced by SARS-CoV-2 (MOI = 0.5) infection for 24 h with different treatments, and the genes belonging to GO annotations for cytokine activity and chemokine activity (GO: 0005125 and GO: 0008009). In c, all error bars, mean values ± SD of five images from n = 5 independent biological mice for each group. P-values are indicated by two-tailed unpaired Student’s t-test. In e, g, all error bars, mean values ± SEM, P-values are determined by unpaired two-tailed Student’s t-test of n = 3 independent biological experiments. For a, b, d, f, similar results are obtained by three independent biological experiments. Source data are provided as a Source data file.

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