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

FIGURE 8.

FIGURE 8

NP and NS1 protein levels and NP mRNA level significantly attenuated post-FLNA over-expression in an IAV microenvironment. (A) HEK293 cells were transfected with pEGFP-N1 or FLNA-GFP plasmid (3 μg) 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), GFP, NP, and NS1. (B,C) Densitometric analysis was performed for the infected, FLNA over-expressed samples using the ImageJ software to visualize FLNA and NP and NS1 protein levels. Results are shown as mean ± S.D. of one independent experiment (n = 3) (see Supplementary Figure 9). Statistical significance for FLNA and NP was determined using one-way ANOVA with post-hoc Tukey test and Student’s t test, respectively (∗p < 0.05). (D) HEK293 cells were transfected with pEGFP-N1 or FLNA-GFP plasmid (3 μg) 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 GAPDH (loading control) mRNA estimation via qRT-PCR. Results are shown as mean ± S.D. of three independent experiments (n = 9). Statistical significance for FLNA and NP was determined using Student’s t test (∗p < 0.05).