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. 2021 Jan 14;10:593805. doi: 10.3389/fcimb.2020.593805

Figure 4.

Figure 4

DRP1 regulates TNF-α production at post-transcriptional level. (A) Quantitative real-time PCR of Tnfa transcript levels of NT-Control and DRP1 KD macrophages after 4h stimulation with LPS or MRSA infection (MOI 20). Data are relative to NT-Control macrophage untreated (Mock) condition. (B) TNF-α levels in culture media of conditions from (A) were quantified by ELISA at 24h post-stimulation. (C) Total TNF-α levels were monitored by intracellular staining using flow cytometry after blocking protein transport with monensin. Macrophages were stimulated with LPS (200 ng/ml), infected with MRSA (MOI 20) or left untreated (Mock) in the presence of monensin 4h and stained intracellularly for TNF-α. Mean fluorescence intensity (MFI) of TNF-α was determined using FlowJo software, representing the geometric mean. Graphs show the mean ± SD from at least three independent experiments. P value was calculated using two-way ANOVA with Sidak’s post-test for multiple comparisons. ***P < 0.001; ****P < 0.0001.