Fig. 6
From: Critical role for arginase 2 in obesity-associated pancreatic cancer
AKT activation enhances in vivo PDA growth and sensitizes tumors to ARG2 knockdown. a Immunoblots for pT308-AKT, pS473-AKT and total AKT in isogenic AsPC1-CFP and AsPC1-AKT PDA cells expressing SRC myristoylation sequence-AKT fusion (myrAKT). b Volumes of orthotopic PDA tumors from cells in a (105 per mouse) grown for 6 weeks in lean (n = 14) or obese (n = 15) 12–14 week old male Rag1 −/− mice following the outline in (Fig. 1a). c, d Immunoblots for pT308-AKT, pS473-AKT, and total AKT c, ARG2 and ARG1 d in tumors from b. e ARG2 levels as quantified by ImageJ and normalized to GAPDH, from immunoblots in d, Fig. 1e, and Supplementary Fig. 10c. f Immunoblots of ARG2 and ARG1 in isogenic cells from a, harboring control Scramble (shScr) or ARG2 (shARG2) knockdown. g Proliferation curves of cells in f (n = 6). Data are representative of four independent experiments. h Volumes of orthotopic PDA xenografts from cells in f that were injected in lean 8–9 week old male Rag1 −/− mice (500,000 per mouse) and grown for 5 weeks (n = 6). i Immunoblots for ARG2, ARG1, pT308-AKT, pS473-AKT and total AKT from representative tumors in h. j, k Representative immunohistochemistry staining showing enhanced ARG2 levels in PDA tumors from patients with higher compared to lower BMI j, and overlapping pS473-AKT and ARG2 staining in sequential tumor sections from PDA patients k. Arrows indicate areas of ductal adenocarcinoma and arrowheads indicate stroma. Scale bars: 20 µm. l Multivariate regression analysis of the association of BMI and pAKT with high ARG2 levels in resected PDA immunostained in j, k. OR odds ratio, CI confidence interval, AJCC American Joint Committee on Cancer. Data represent the mean ± s.d. in b, e, h or mean ± s.e.m. in g. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, one-way ANOVA followed by Tukey test
