Fig. 2

rHcADRM1 proteins suppressed cell viability and induced apoptosis via the interaction with goat T cells. a Determination of the interaction of HcADRM1 protein with goat T cells in vitro. T cells treated with (a₁) or without (a₂) rHcADRM1 protein were incubated with rat anti-rHcADRM1 IgG as the primary antibody. T cells stimulated by rHcADRM1 were incubated with normal rat IgG as the primary antibody (a₃). b rHcADRM1 significantly inhibited T cell viability. Cell viability was determined via the incorporation with CCK-8, whereas cell viability index was determined by calculating the OD 450 values of the control group as 100%. Results are presented as the mean ± SD. Asterisks denote statistically significant differences (*P < 0.05, ***P < 0.001, ****P < 0.0001) compared with the control group. c Flow cytometry analysis of T cell apoptosis in responses to rHcADRM1 stimuli. Apoptosis determination was performed via 7-AAD and Annexin V-PE staining. d Statistical analysis of T cell apoptosis. The apoptotic proportions were counted via the AnnexinV⁺7AAD⁻ and AnnexinV⁺7AAD⁺ T cells. Data are presented as the mean ± SD, where an asterisk denotes a statistically significant difference (*P < 0.05, ****P < 0.0001) compared with the control group. Each experiment was performed in triplicate. e The mRNA transcripts of caspase-8, -9 and -3 in rHcADRM1-stimulated T cells. Results are denoted as minimum to maximum (all points). Asterisks denote statistically significant differences (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001) compared with the control group. Each experiment was run in triplicate. Scale-bars: a 100 µm