ELISA measurement of IL-6 and IL-10 levels

The cell culture medium and treated tissue samples were collected and centrifuged (1,000 ×g) for 5 min. After the centrifugation, the cell-free supernatant was obtained. The solution was blocked by PBS for 1 h and was added to 100 µL of supernatant followed by 2 h of incubation at room temperature. Subsequently, 100 µL anti-mouse IgG antibody was added and incubated at room temperature for a further 2 h. Next, 100 µL of TMB substrate was added, and the reaction was terminated after 30 min of dark color development at room temperature. The enzyme reader determined the OD value of the sample at 450 nm, and the sample concentration was calculated.