Hmga2 overexpression alters gene expression in Jak2^V617FLSK. (A) Heat maps showing significantly upregulated and downregulated genes in Jak2^VF/+-Hmga2 Lin^–Sca-1⁺ckit⁺ (LSK) cells compared with Jak2^VF/+-vector LSK cells. (B) A list of selected genes that are significantly upregulated in Jak2^VF/+-Hmga2 LSK cells compared with Jak2^VF/+-vector LSK cells. (C) Gene set enrichment analyses (GSEAs) of the RNA-seq data from Jak2^VF/+-vector LSK cells and Jak2^VF/+-Hmga2 LSK cells. Enrichment plots of selected gene sets with normalized enrichment score (NES) and false discovery rate (FDR) are shown. (D) Relative expression of Cxcl12, Fzd2, collagen1a1 (Col1a1), collagen3a1 (Col3a1), Ifi27l2a, Gata3, Dok4, Meis1, and Tgf-βR2 mRNA was determined in Jak2^VF/+-vector and Jak2^VF/+-Hmga2 LSK cells by RT-qPCR and normalized with glyceraldehyde 3-phosphate dehydrogenase (Gapdh) expression. (E) Validation of some HMGA2 targets in JAK2^V617F-positive human megakaryoblastic SET-2 cells. SET-2 cells were transduced with lentivirus expressing HMGA2, and the infected cells were selected using puromycin. Relative expression of CXCL12, FZD2, IFI27 (homolog of mouse Ifi27l2a), GATA3, DOK4, MEIS1, and TGF-βR2 was assessed by RT-qPCR and normalized by GAPDH. Data from 4 independent experiments are shown in bar graphs as mean ± SEM. (F) Validation of the target genes by knockdown of HMGA2 in SET-2 cells. SET-2 cells were transduced with lentiviral HMGA2 short hairpin RNA (shRNA) or scramble shRNA (control), and the infected cells were selected by using puromycin. Relative expression of CXCL12, FZD2, IFI27, GATA3, DOK4, MEIS1, and TGF-βR2 was assessed by RT-qPCR and normalized by GAPDH. Data from 4 independent experiments are shown in bar graphs as mean ± SEM. *P < .05.