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. 2018 Apr 20;13(4):e0192709. doi: 10.1371/journal.pone.0192709

Fig 2. RSV induced IL-1β regulates OPN expression during RSV infection.

Fig 2

(A-B) HEK-293 cells were mock-infected or infected with 1 MOI of RSV-L19F. 2 hours after the infection cells were treated with increasing concentrations of human rIL-1β (0, 1 or 10 ng/ml). (A) Protein was harvested 24 hpi for western blots. 25μg of protein lysate was loaded in each lane. OPN is seen at 55 kDa and β-actin (loading control) at 42 kDa. (B) Plaque viral titers were obtained from the supernatants collected 24 hpi. (C-E) HEp-2 cells were pre-treated two hours prior to RSV-L19F infection with 10 μM of Ac-YVAD-CHO (caspase-I) inhibitor. Cells were infected with 1 MOI of RSV-L19F and infectious media was replaced with fresh media containing 10 μM of Ac-YVAD-CHO. RNA and protein were collected 24 hpi. (C and D) Expression levels of IL-1β and RSV-N were determined by qPCR. Results are presented as fold-change in expression of IL-1β or RSV-N mRNA normalized to the control (HPRT). (E) 25μg of protein lysate was loaded in each lane. OPN is seen at 55 kDa and β-actin (loading control) at 42 kDa. qPCR data are represented as means ±SEM. Experiments were performed in triplicate. **** p < 0.0001.