NSP1 downregulates steady-state levels of mRNAs encoded by immune-related genes.

A.) A549 cells and their growth medium were collected 16 hours after mock, NSP1-GFP or FH-NSP1 transfection. Growth media and cell extracts were analyzed by western blot, and samples from FH-NSP1 cells were also used for immunoprecipitation with anti-FLAG antibodies prior to Western blot analysis using an anti-NSP1 antibody. B.) Confocal fluorescence microscopy of A549 cells transfected with NSP1-GFP or mock (same transfection and processing in the absence of DNA). C,D) Principal Component Analysis (PCA) of mRNA profiles after transient transfection of NSP1 and control GFP or the K164A/H165A NSP1 mutant, obtained by RNA-seq following rRNA depletion (C) or poly-A selection (D). E.) Gene ontology analysis of transcripts downregulated (≥2-fold) upon expression of NSP1. Size of bubble reflects the number of downregulated genes of the respective categories. F.) Venn diagram of genes downregulated by ≥ 2-fold upon NSP1 expression compared to expression of the GFP control, using either rRNA depletion or poly-A selection for mRNA enrichment. Genes shared by the two groups are listed below.