BJ-3105 slightly inhibits CD4<sup>+</sup> T cell proliferation without inducing apoptosis.

<p>CFSE-labeled naïve CD4<sup>+</sup> T cells were activated with anti-CD3 anti-CD28 under the treatment of indicated dose of BJ-3105 for 3 days in (A) Th1 and (B) Th17 differentiation conditions. Cell proliferation was monitored by assessing CFSE dilution by flow cytometry. (C) Naïve CD4<sup>+</sup> T cells and APCs from spleens and lymph nodes were isolated and activated under Th1 differentiation condition in the presence of BrdU (10 μM) with or without BJ-3105 (1 μM) and analyzed after 3 days by flow cytometer. Representative bar diagram for percentage of BrdU<sup>+</sup>CD4<sup>+</sup> T cells were shown. (D) Representative Dot plots examining CD4<sup>+</sup> T cell proliferation by detecting Ki-67 in Th1 differentiation condition with or without BJ-3105 by flow cytometer. Percentage of Ki-67 positive CD4<sup>+</sup> T cells were shown. (E) Live cells were detected by Anexin-V and PI staining after cultured in Th1 differentiation conditions for 72 h and analyzed by flow cytometer. (F) Percentage of live cells on dose dependent treatment of BJ-3105. Representative results of three experiments are shown. <i>*p</i> < 0.05, compared with drug untreated group. Data shown are mean ± SEM.</p>