Fig. 2

Agonistic activity and MPL signal transduction in mouse and human cells by 2R13. a-c Cell proliferation assay for detecting the agonistic activity of 2R13. a BaF3/MPL cells were incubated with rhTPO (positive control) or 2R13, and b Parental BaF3 cells were incubated with WEHI-CM (positive control), rhTPO, or 2R13, over the indicated concentration range (n = 3). The EC₅₀ was calculated by fitting the data to a four-parameter logistic model using GraphPad Prism 9. c M07e cells were incubated with rhTPO (positive control) or 2R13 over the indicated concentration range (n = 3). d Western blotting of BaF3/MPL cells stimulated with rhTPO or 2R13 over the indicated concentration range for 30 min, using the indicated antibodies. e Luciferase reporter assay in BaF3/MPL cells transfected with pGL4.52 [luc2P/Stat5RE/Hygro] plasmids. Cells were stimulated with rhTPO (positive control), control antibody (negative control), or 2R13 over the indicated concentration range for 6 h (n = 2). Data are the mean ± SD. WEHI-CM, WEHI-3B cell-conditioned medium