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. 2020 Sep 25;11:581867. doi: 10.3389/fmicb.2020.581867

FIGURE 7.

FIGURE 7

NP and NS1 protein levels and NP mRNA level significantly increased post-FLNA silencing in an IAV microenvironment. (A) A549 cells were transfected with FLNA (200 nM) or non-targeting control (CTRL) siRNA (200 nM) for 24 h followed by IAV PR8 infection (MOI = 5). 24 h.p.i the cells were harvested with RIPA buffer and 30 μg of protein lysate was loaded onto an SDS-PAGE gel followed by Western blotting. Blots were developed by ECL for FLNA, vinculin (loading control), NP and NS1. (B,C) Densitometric analysis was performed for the infected, FLNA silenced samples using the ImageJ software to visualize FLNA and NP and NS1 protein levels. The data show mean ± S.D. from one representative experiment (n = 3) (see Supplementary Figure 7). Statistical significance was determined using one-way ANOVA with post-hoc Tukey test and Student’s t test, respectively (∗p < 0.05). (D) A549 cells were transfected with FLNA (200 nM) or non-targeting control (NT) siRNA (200 nM) for 24 h followed by IAV PR8 infection (MOI = 5). 24 h.p.i the cells were harvested and total RNA was extracted followed by NP and β-actin (loading control) mRNA estimation via qRT-PCR. Results are shown as mean ± S.D. of three independent experiments (n = 9). Statistical significance was determined using Student’s t test (∗p < 0.05).