Biomolecules & Therapeutics : eISSN 2005-4483 / pISSN 1976-9148

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Fig. 4. Effects of pathway-specific inhibitors on SARS-CoV-2-induced STAT1 and STAT3 phosphorylation in Calu-3 cells. (A) Calu-3 cells were pretreated with 0.1% DMSO, 10 μM PD169316, 25 μM PD98059, 25 μM SP600125, 25 μM AG490, or 1 μM JAK inhibitor I for 30 min. (B) Calu-3 cells were pretreated with the indicated concentrations of S3I-201 inhibitor for 30 min. The cells were washed with PBS and then infected with SARS-CoV-2 in PBS at 0.5 MOI. After 1 h of incubation, the medium was replenished with DMEM containing 2% FBS. (C) Calu-3 cells were treated with 0.1% DMSO, 10 μM PD169316, 25 μM PD98059, 25 μM SP600125, 25 μM AG490, 1 μM JAK inhibitor I, or 20 μM S3I-201. (A-C) After 48 h of incubation, cell lysates were prepared, and Western blotting was performed with the indicated antibodies. (D) Calu-3 cells were pretreated with 0.1% DMSO, 25 μM AG490, 1 μM JAK inhibitor I, 20 μM S3I-201, or 10 μM STA-21 for 30 min. The cells were washed with PBS and then infected with SARS-CoV-2 in PBS at 0.5 MOI. After 1 h of incubation, the medium was replenished with DMEM containing 2% FBS. After 48 h of incubation, the cells were treated with 20 nM Leptomycin B for 3 h. To evaluate phosphorylation and localization of STAT3, cells were fixed with 4% paraformaldehyde, stained with phospho-STAT3 antibody (green), and observed by confocal microscopy. Hoechst 33258 was used for nuclei staining (blue).
Biomolecules & Therapeutics 2021;29:273~281 https://doi.org/10.4062/biomolther.2020.226
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