Figure 7. TGF-β1 expression is increased in patient derived MPN samples.

(A) Gating strategy for TGF-β and MPL quantification on CALR^del52 or CALR^WT iPSC-derived erythroid progenitor cells CD45⁺CD71⁺CD235a^+/-.

(B) Representative histogram plot showing TGF-β expression on the surface of CALR^del52 or CALR^WT iPSC-derived CD45⁺CD71⁺CD235⁺ erythroid progenitors with respective unstained control.

(C) Scatter plot showing relative MFI of L-TGF-β surface expression on CALR^WT or CALR^DEL52 iPSC-derived CD45⁺CD71⁺CD235a⁺ erythroid progenitor cells, fold change to MFI of unstained control. CALR^DEL52 iPSC and CRISPR repaired CALR^WT control were generated from one CALR^del52 MPN patient. Each dot represents individual experiment. The P-value was calculated using unpaired Student´s t test.

(D) Scatter plot showing fold change RNA expression of human TGF-β1 in total mononuclear BM cells from MPN patients or healthy donors from the Freiburg MPN cohort.

(E) Box plot showing TGF-β1 RNA expression (log2) in CD34⁺BMMCs/PBMCs from publicly available data set (GSE174060 - (42)).

(F) Heat map showing expression profiling by GeneChipHuman Gene 2.0 ST Arrays (Affymetrix) of Lin^-CD34⁺ PBMCs of normal bone marrow donors (NBM), patients with primary myelofibrosis (MF) and secondary AML (sAML). Data were obtained and re-analyzed from Gene Expression Omnibus (GEO) Database - GSE214361 (43). A set of genes involved in TGF-β1 regulation and activation was selected. Z score for separate patients is shown on the left and fold change comparing each group is shown on the right (* indicates significant changes).