Figure 4. DZNep treatment modulates tissue and immune processes.

• A–D
  Mass spectrometry‐based analysis of cells treated with DZNep and infected with SARS‐CoV‐2 and SARS‐CoV. (A) Schematic representation of LC–MS/MS experiments. A549‐ACE2s and NHBEs were pretreated for 6 h with 0.75 and 1.5 μM DZNep, respectively, or vehicle (PBS), and infected with SARS‐CoV‐2 or SARS‐CoV at MOI 3 for 24 h (A549‐ACE2) or 36 h (NHBEs). Changes in protein abundance were analyzed according to the depicted scheme using LASSO‐based linear model followed by fixed LASSO inference‐based p‐value estimation as described in the Materials and Methods section. (B) Number of significantly up‐ or downregulated proteins in indicated comparisons according to (A). (C) Donor‐normalized LFQ abundance of viral nucleoprotein (N) and spike (S) in the indicated conditions. Error bars represent mean ± SD of n = 4 donors (NHBE) or n = 4 independently infected A549‐ACE2 cultures. Statistics were calculated using Student's two‐sided t‐test as indicated. (D) Expression patterns according to (A) of a selection of genes related to the disease‐relevant pathways as annotated.
• E
  NHBEs (six independent donors) were pretreated for 6 h with 0.75 μM DZNep or vehicle and infected with SARS‐CoV or SARS‐CoV‐2 at MOI 3. Twenty‐four hours later, accumulation of IL‐6 and IP‐10 was measured in the supernatant by ELISA. Donor‐wise IL‐6 and IP‐10 secretion, normalized to vehicle‐treated uninfected controls (as further described in Materials and Methods), is shown.
• F
  Schematic representation of the proposed disease‐relevant functions of DZNep in the context of COVID‐19 alongside the model of their molecular origin.