Fig. 5: Overexpression of HMGA2 in classical PDAC results in more aggressive disease and higher levels of pS6K in vivo.
From: HMGA2 and protein leucine methylation drive pancreatic cancer lineage plasticity
a WB for pS6K T389, total S6K, pEIF4B S422, total EIF4B, pS6 S235/S236, total S6, and His-tagged HMGA2 in three classical PDAC cell lines engineered to overexpress empty vector or HMGA2 WT upon doxycycline treatment. b WB of puromycin incorporation, LIN28B, ACTIN, His-tagged HMGA2, and H3 in four HMGA2low PDAC cell lines engineered to overexpress empty or HMGA2 WT upon doxycycline treatment. WCL, whole cell lysate; Chr, chromatin. c Engineered Capan-2 and HPAF-II cells outgrow control cells when overexpressing WT HMGA2 (red). Subcutaneous flank implants in NSG mice, treated with 200 ug/mL doxycycline post-surgery. Data are displayed as means ± s.e.m. N(Capan-2 Empty) = 4 mice, N(Capan-2 WT HMGA2) = 4 mice, N(HPAF-II Empty) = 4 mice and N(HPAF-II WT HMGA2) = 4 mice; P values after paired, two-tailed t test. d Tumor-free survival curve for mice orthotopically implanted with the indicated engineered Capan-2 cell lines. N(Capan-2 Empty) = 6 mice and n(Capan-2 WT HMGA2) = 6 mice; P values after log-rank test. NSG mice were treated with 200 µg/mL doxycycline post-surgery. e Metastases to the lung (left, scale bar = 50 µm) and small intestine (right, scale bar = 100 µm) were observed with HMGA2-overexpressing tumors. Specific immunohistochemistry for HMGA2. Scale bar = 50 µm. f Quantification of specific immunohistochemistry for HMGA2 and pS6K T389 in HMGA2 WT tumors compared to empty vector control of orthotopically implanted engineered Capan-2 cells from Fig. 5d. Box plots represent median, interquartile range and 5–95th percentile of n = 6 mice. Asterisks, P values after unpaired, two-tailed t test. **P = 0.005 HMGA2 WT vs EMPTY, pS6K; ****P < 0.0001 HMGA2 WT vs EMPTY, HMGA2. g Representative HMGA2 and pS6K IHC images. Scale bar = 50 µm. Source data are provided as a Source Data file.
