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Fig. 5 | Gut Pathogens

Fig. 5

From: Translocating lipopolysaccharide correlates with the severity of enterovirus A71-induced HFMD by promoting pro-inflammation and viral IRES activity

Fig. 5

The effect of LPS post-treatment on proliferation of EV-A71 in vitro. RD cells and SH-SY5Y cells were infected with BrCr Strain EV-A71and neurotropic Strain EV-A71 at an MOI of 2, respectively. 2 h later, the cells were treated with LPS at the concentration of 200 ng/mL or 500 ng/mL, respectively. At 12 h post infection, photomicrographs were taken (original magnification, ×100) (a), the protein levels of viral VP1 in cell lysates were measured by western blot (b). RD cells or SH-SY5Y cells were pre-transfected with p-EGFP-Vector (Vec, 2 μg/well, 6-well plate) or p-EGFP-2A (2A, 2 μg/well, 6-well plate), respectively. Subsequently, 12 h later, the cells were re-transfected with Cap-Rluc-vIRES-Fluc mRNA (100 ng/well, 96-well plate). 4 h later, the cells were treated with 200 ng/mL or 500 ng/mL LPS for another 12 h. The intensities of Fluc and Rluc were detected as described in MMs. The results (Rluc/Fluc) indicate the M ± SD of three independent experiments (c, d). RD cells or SH-SY5Y cells were pre-transfected with p-EGFP-Vector (Vec) or p-EGFP-2A (2A), respectively. 12 h later, the cells were treated with 500 ng/mL LPS for another 12 h. The cell lysates were used for the protein detection of eIF4GI, phosphorylated-ERK (p-ERK) and total ERK (t-ERK) by western blot (e). EV, EV-A71; EVL200, EV-A71 + 200 ng/mL LPS; EVL500, EV-A71 + 500 ng/mL LPS. 2AL200, 2A + 200 ng/mL LPS; 2AL500, 2A + 500 ng/mL LPS. Statistical difference was determined by student’s t test. n.s. > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001

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