Lps tlrl peklps

We used MT-iPS-MLs and homogeneous time-resolved fluorescence (HTRF)-based chemokine measurements for the HTS system. The compound library is described in Table 1. For the 1st and 2nd screenings, 5ൈ10 3 MT-iPS-MLs were seeded into each well of 384-well plates. The plates were individually prepared for the detection of MCP-1 or IP-10. The cells were treated with compounds at concentrations of 1 μM (1st screening) and 100 nM (2nd screening). Three hours later, 50 ng/mL lipopolysaccharide (LPS) (tlrl-peklps; InvivoGen) for MCP-1 and 100 ng/mL IFN-γ for IP-10 was added for the stimulation. After the subsequent culture for 21 hours, the supernatants were sampled, and the Human CCL2 (MCP-1) Kit (62HCCL2PEG; Cisbio) or Human CXCL10 (IP-10) Kit (62HCX10PEG; Cisbio) was added to detect MCP-1 or IP-10 (Fig. 1A). The chemokine concentration was detected by using POWERSCAN ○ R 4 (DS Pharma Biomedical). Validation of the hit compounds with MT-ES-MLs was performed using the same procedure as the 2nd screening.