Journal: bioRxiv

Article Title: The cold-inducible RNA-binding protein RBM3 stabilises viral mRNA at the cooler temperatures of the upper respiratory tract

doi: 10.1101/2024.07.25.605104

Figure Lengend Snippet: (A) Cold-responsiveness of a panel of cell lines and primary nasal epithelial cells (PNECs) as determined by the induction of RBM3 expression by qPCR over 48 h and normalised to 37°C levels. (B) Generation of a lentiviral RBM3 overexpression (RBM3+) A549 cell line alongside a control empty vector line (EMP). (C) Titres of WSN at 72 hpi after an MOI 0.01 infection in EMP or RBM3+ A549 cells. (D) RBPmap predicted RBM3 binding motifs for each WSN positive strand segment, separated by predicted confidence of each motif. (E) Western blot showing input and pulldown samples for CLIP-qPCR, blotting for Actin and overexpressed RBM3-Flag. (F) qPCR quantification of the RNA extracted from CLIP samples demonstrated a significant increase in viral NP mRNA levels with minimal differences in HA or Actin transcripts, when comparing RBM3 to no antibody control pulldowns.

Article Snippet: Cells were submerged in modified Promocell Airway Epithelial Cell Growth Medium which was replaced on the apical and basal side every 2 days until the cells reached confluency.

Techniques: Expressing, Over Expression, Control, Plasmid Preparation, Infection, Binding Assay, Western Blot