Journal: Communications Biology

Article Title: NRF2 activators and the inhibitor of nuclear export, selinexor, restrict coronaviruses by targeting a network involving ACE2, TMPRSS2, and XPO1 through an NRF2-independent mechanism

doi: 10.1038/s42003-026-09724-6

Figure Lengend Snippet: A –C Reduction of ACE2 , TMPRSS2 , and XPO1 mRNA. RT-qPCR analysis of RNA from the experiment shown in Fig. . D 4OI and SEL reduce ACE2 and TMPRSS2 protein levels. Uninfected Calu-3 cells were grown in medium containing 4OI (100 µM) or SEL (1 µM), and cellular ACE2 and TMPRSS2 levels were measured by immunoblot after 1–72 h. E 4OI reduces half-life of ACE2. Uninfected Calu-3 cells were grown in medium containing 4OI with or without cycloheximide (CHX, 50 µg/ml), and cellular ACE2 levels were measured by immunoblot after 1–12 h. Densitometry of the blots shown in ( E ); F , bar chart; G , numerical values; untreated cells = 100%. H NEDD4L knock-down attenuates the ACE2-destroying capacity of 4OI at the protein. Efficiency of NEDD4L mRNA knock-down is shown in Supplementary Figure . I . MDM2 knock-down attenuates the ACE2-destroying capacity of 4OI at the protein level. Efficiency of MDM2 mRNA knock-down is shown in Supplementary Figure . J –N Uninfected Calu-3 cells were grown in medium containing 4OI, SEL, proteasome inhibitor (CFZ, 10 nM), or lysosome inhibitors (BAFA1- 100 nM, CQ- 50 µM), and expression of ACE2 was determined by immunoblot. CFZ 6 h ( J ), BAFA1 24 h ( K , L ), and CQ 24 h ( M , N ). O 4OI and SEL reduce XPO1 protein levels. Uninfected Calu-3 cells were grown in medium containing 4OI or SEL, and XPO1 levels were measured by immunoblot after 1–72 h. P , Q NEDD4L and MDM2 knock-down attenuate ACE2 mRNA reduction by 4OI and SEL (RT-qPCR after 24 h). R 4OI and SEL attenuate STAT3 phosphorylation (immunoblot after 24 h). n = 3 ( D , E , H , I , O : n = 2), means ± SEM. One-way ANOVA with Tukey’s post-hoc test. * ≤0.05, ** ≤0.01, *** ≤0.001, **** ≤0.0001.

Article Snippet: The following primary antibodies were used: ACE2 (Cell Signaling Technology, 92485S, 1:1000), XPO1 (Cell Signaling Technology, 46249S, 1:1000), TMPRSS2 (Santa Cruz, sc-515727, 1:500), MDM2 (Cell Signaling Technology, 86934S, 1:1000), NEDD4L (ThermoFisher Scientific, 67276-1-IG-20UL, 1:10,000), P-STAT3 (Cell Signaling Technology, 9145S, 1:1000), STAT3 (Cell Signaling Technology, 4904S, 1:1000), and β-actin (Abcam, ab49900, 1:20,000).

Techniques: Quantitative RT-PCR, Western Blot, Knockdown, Expressing, Phospho-proteomics