TA-316 induces maturation of imMKCLs and increases functional platelets. imMKCLs cultured with rhTPO (50 ng/mL), TA-316 (200 ng/mL), eltrombopag (1000 ng/mL), or DMSO (0.05%) were collected on day 11 (genes on) and day 15 (after 4 days without DOX, genes off). (A) Multinucleated (≥8 n) MKs and (B) PAC-1 binding on platelets were analyzed by FACS. Platelets were stimulated for 15 minutes with 1 U/mL thrombin from human plasma. Human washed platelets served as a positive control. (C) mRNA expressions of GATA1, ZFPM1 (FOG1), NFE2, FLI1, and VWF (genes on or off) were evaluated using Taqman PCR assays. GAPDH served as an internal control. Data represent the mean and SD; *P < .05, **P < .01. PLT, platelets.