Fig. 7: STAP-1 regulates STAT5 via JAK and PPAR signaling in CML.

a STAP-1 knockdown CML cell lines were established using short hairpin RNA. Total RNA samples isolated from these cells were quantified by real-time RT-PCR. Data represent the levels of these mRNAs normalized to control (sh control). Data of a representative experiment, which was repeated at least three times with similar results, are shown. b Phosphorylation status of STAT5 (pY649) in CML cell lines was analyzed by flow cytometry. Bar graph represents MFI of phosphorylated STAT5 normalized to sh control cells (n = 6). CML cell lines were treated with imatinib (c) or ruxolitinib (d) at the indicated concentrations for 24 h. Growth inhibitory effect on these cells was determined. The data represent the means of triplicate experiments. Similar results were obtained for three independent experiments. e 293T cells were transfected with STAT5a, STAT5b, or JAK2 in either presence or absence of Myc-tagged STAP-1. The cells were lysed and immunoprecipitated with anti-Myc antibody. An aliquot of total cell lysate (TCL) and the immunoprecipitates (IP) were blotted with anti-STAT5a, anti-STAT5b, anti-JAK2, anti-Myc, or anti-STAP-1 antibody. f STAP-1 knockdown CML cell lines were either or not treated with pioglitazone at 50 μM for 96 h and phosphorylation status of STAT5 (pY649) was analyzed. Data of a representative experiment, which was repeated at least three times with similar results, are shown. Data represent the mean ± SD. *p < 0.05, **p < 0.01, and ***p < 0.001 indicate significant difference.