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. 2019 Oct 25;10:2438. doi: 10.3389/fmicb.2019.02438

FIGURE 7.

FIGURE 7

Effect of LCMV infection on HIF-1 and PI3K/Akt activation. (A) Mock- and LCMV-infected HeLa cells were transfected with HRE-Luc, and the reporter protein (luciferase) activity was determined at 48 h post-transfection. Reporter protein activity was normalized to uninfected, control-transfected cells. Values represent means of three independent experiments, each done in eight replicates. Error bars denote standard deviations. Data are presented as a relative increase compared to control (HeLa cells), which was set to 1. ****P < 0.0001 (HeLa-MX vs. HeLa). (B) Immunoblot analysis of p-Akt (S473), total Akt, and viral NP with specific antibodies using whole-cell extracts prepared from untreated (−) mock-infected HeLa cells (HeLa) and LCMV-infected HeLa cells (HeLa-MX) or cells treated (+) with 10 mM NAC for 24 h. The signal obtained with anti-β-actin antibody was used as loading control. One representative of at least four biological replicates is shown. (C) Densitometric analysis of protein abundance was performed using ImageJ software or Image Studio Lite software. Relative quantity of p-Akt was calculated as the ratio of the intensity of each signal to the intensity of the related signal for total Akt. Values represent the means of six biological replicates. Error bars denote the standard deviations. Data are presented as relative change compared to control (untreated HeLa cells), which was set to 1. ∗P = 0.0119 (HeLa vs. HeLa-MX), ∗P = 0.0101 (HeLa-MX vs. HeLa-MX NAC), P = 0.3002 (HeLa vs. HeLa NAC). (D) Densitometric analysis of protein abundance was performed using ImageJ software or Image Studio Lite software. Relative quantity of NP was calculated as the ratio of the intensity of each signal to the intensity of the related β-actin or tubulin internal standard, respectively. Values represent the means of four biological replicates. Error bars denote the standard deviations. Data are presented as relative change compared to untreated HeLa-MX cells, which was set to 1. ∗∗P = 0.0059 (HeLa-MX NAC vs. HeLa-MX).