Thermal stability of Mpro in the presence or absence of GRL-0920. Thermal stability of Mpro in the presence of GRL-0920 was analyzed using differential scanning fluorimetry (DSF) as described for experiment 1. Note that the Tm value of Mpro (5 μM) alone was 53.63°C, while the Tm values decreased to 51.04, 49.13, and 46.98°C in the presence of 5, 50, and 100 μM GRL-0920, respectively, suggesting that the Mpro get destabilized when GRL-0920 covalently bound to Mpro. Note that, as expected, the presence of HIV-1 protease inhibitor GRL-142 (5 μM) increased the thermal stability of HIV-1 protease (5 μM) as shown for experiment 2.