Figure 2
Adipocytes sustain pancreatic preneoplastic lesions aggressiveness through a WNT paracrine network. (a) Interaction network derived from secreted genes upregulated in adipocytes versus the hBMSC cells from two healthy donors. Each interaction is supported by at least one literature reference identified in the Ingenuity Pathway Knowledge Base, with solid lines representing direct interactions and dashed lines representing indirect interactions. In the heat map shown in this figure, the logarithms of the gene expression levels are shown in colors (green=decreased expression, red=increased expression). (b) Quantitative real-time PCR gene expression analysis of WNT5A, WNT5B, RSPO1, DKK1, DKK2, DKK3, SFRP2 and FRZB in hBMSC and relative adipocytes from two healthy donor (#1 and #2). The result values represent the values of the fold change in RNA expression between the gene of interest and b-actin. The mean values and 95% CI from three independent experiments done in quadruplicate are shown. ***P<0.001, **P<0.005 and *P<0.05, by two-tailed unpaired Student’s t-tests. (c, d) Western blot analysis for GSK3β and phospho-GSK3 β in HPDE and HPDE/KRAS cells cultured with conditional medium (CM) from hBMSC (hBMSCCM) or adipocytes (ADIPOCM) from two healthy donor. γ-Tubulin was used as loading control. The relative protein quantification, using ImageJ software (https://imagej.nih.gov/ij/), was performed and relative GSK3 and γ-tubulin ratio are indicated.
