Fig 8. IL-6 blockade attenuates astrocytic cell apoptosis upon EV71 infection in U251 cells.

(A) The schematic of IL-6 antibody neutralization upon EV71 infection. 2×10⁶ U251 cells were blocked with neutralized mouse anti-human IL-6 antibody (0.5 μg/ml) or mouse isotype IgG (0.5 μg/ml) at 2 h post EV71 infection (MOI = 0.5). (B) The supernatants of treated cells were collected and IL-6 protein level was measured by ELISA. (C and D) Cell viability (C) and LDH release (D) of treated cells for 12, 24 or 48 h were examined using CCK8 and LDH assay, respectively. Data are shown as mean ± SD and correspond to a representative experiment out of three performed. ns, non-significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001. (E) The RNA was extracted from the transfected cells at 12 or 24 h post EV71 infection. The levels of apoptosis-associated genes mRNAs were measured by qPCR. Data are shown as fold changes of RNA expression compared to mock samples. (F) The protein was extracted from the transfected cells at 24 h post EV71 infection and then detected by Western blotting with targeted antibodies. Relative PARP protein expression to internal control is quantified using Image J software.