Fig 2: Bazedoxifene inhibits signal transducer and activator of transcription (STAT)3 phosphorylation and expression of STAT3 downstream genes, and induces apoptosis. HEPG2 (A), 7721 (B) and HUH-7 (C) cancer cell lines exhibit constitutively phosphorylated STAT3. Bazedoxifene (10, 15, 20, 25 µmol/L) inhibited the phosphorylation of STAT3 (Tyr 705), Janus kinase (JAK)1 and JAK2, induced apoptosis as evidenced by increased cleaved caspase-3. The expression of STAT3-mediated genes was decreased by bazedoxifene except for the expression of Bcl-2 in 7721 cells. GAPDH, glyceraldehyde 3-phosphate dehydrogenase

Fig 3: Small-scale mutations in genes of the JAK-STAT pathway are predominant in pcAECyTCL. (A) Diagrams showing deleterious indels and single nucleotide variants (SNV) in JAK2, JAK3, STAT3, STAT5B, SH2B3 and SOCS1 detected in primary cutaneous CD8+ aggressive epidermotropic cytotoxic T-cell lymphoma (pcAECyTCL) by whole-genome sequencing (Table 1). (B) Sanger chromatograms confirming presence of bona fide pathogenic SNV in patients with pcAECyTCL. (C) Summary of genetic alterations affecting members of the JAK-STAT pathway in pcAECyTCL.

Fig 4: Crlf2 and mutant Jak2 cooperate to induce murine B-ALL development in vivo. (A) Schematic of the Eµ-Crlf2 transgene. (B) Schematic overview of a fetal liver transplantation mouse model of leukemia. Fetal liver cells isolated at embryonic day 13.5 (E13.5) from Eµ-Crlf2 and wild-type C57BL/6 CD45.2+ mice were transduced with pMSCV-IRES-GFP (MIG), MIG-Jak2WT, MIG-Jak2R683G, or MIG-Jak2P933R and transplanted into sublethally irradiated (9.0 Gy) wild-type C57BL/6 CD45.1+ recipients. The resulting primary leukemias were collected and retransplanted into secondary wild-type C57BL/6 CD45.1+ recipients to derive a transplantation model for subsequent functional in vivo experiments. (C) Kapler-Meier survival curves of cohorts of wild-type C57BL/6 CD45.1+ mice transplanted with primary Eµ-Crlf2/Jak2R683G (M#9; n = 8) and Eµ-Crlf2/Jak2P933R (M#8; n = 6) cells. (D) At terminal disease or the conclusion of the experiment, mice in C were autopsied, and splenic tumor burden was assessed by weight. (E) Hematoxylin–eosin (H&E)-stained spleen and liver sections from moribund wild-type recipient mice transplanted with primary Eµ-Crlf2/Jak2R683G and Eµ-Crlf2/Jak2P933R cells. Bars, 100 µm. (F) Light microscopy of May-Grunwald-Giemsa-stained peripheral blood smears from moribund secondary recipient mice of Eµ-Crlf2/Jak2P933R cells show marked numbers of medium to large blast cells with high nuclear to cytoplasmic ratio. (G) Flow cytometry analysis of Crlf2 and GFP expression on splenocytes isolated from C57BL/6 wild-type mice and moribund secondary recipient mice transplanted with Eµ-Crlf2/Jak2R683G cells. A representative flow cytometry plot is shown. (H) Immunoblotting was performed against the indicated targets using lysates isolated from nontransformed C57BL/6 wild-type splenocytes, GFP+ Eµ-Crlf2/Jak2R683G, and Eµ-Crlf2/Jak2P933R cells. Hsp90 served as loading control. (I) Nontransformed C57BL/6 wild-type splenic B-cells, GFP+ Eµ-Crlf2/Jak2R683G, and Eµ-Crlf2/Jak2P933R cells were assayed by PCR of genomic DNA (gDNA) for VDJ rearrangements using the indicated V gene family-specific VH primers. (J) Flow cytometric analysis of the GFP+ Eµ-Crlf2/Jak2R683G cell population reveals a pre-B-cell immunophenotype (B220lowIgM-IgD-CD43-CD24+ckit+CD11b-CD3-).

Fig 5: Cardiac aKO?KO mice exhibit FTT with impaired liver GH signaling ADaily body weight of aKO?KO and littermate control mice. n = 55–145 mice per group with both genders included.BRepresentative picture of 10-day-old aKO?KO and littermate control aHet?WT mice.C, DPlasma GH and IGF1 concentrations in 10- (C, n = 7–10 mice per group) and 16-day-old mice (D, n = 9–11 mice per group) measured by ELISA.EPhosphorylated (Tyr694) and total STAT5 in 10-day-old mouse livers determined by Western blot (n = 3–4 mice per group). ß-Actin serves as a loading control.FRelative levels of phosphorylated and total STAT5 and JAK2 in 13-day-old mouse livers (normalized to total protein content of individual mouse liver) were quantified by ELISA (n = 13 mice per group).GExpression of STAT5 target genes Igf1, Igfbp3, and Igfals in 10-day-old mouse livers measured by qPCR (n = 7–8 mice per group).HLiver (normalized to total protein content, n = 15–16 mice per group) and plasma (n = 9–10 mice per group) IGFBP3 concentrations in 13-day-old mice measured by ELISA.Data information: *P < 0.05, **P < 0.01, ****P < 0.0001, *****P < 0.00001, and ******P < 0.000001 between aKO?KO and all other control genotypes by t-test. All values are mean + s.e.m. Source data are available online for this figure.