Fig. 3: Loss of DUSP5 results in increased levels of activated ERK1/2 and accelerates the transdifferentiation of pancreatic acinar cells in vitro.

A TaqMan RT-qPCR analysis showing mRNA levels of Dusp5, relative to Beta-actin (Actb) following RNA isolation from 3D acinar cultures at 0 and 72 h. Mean values from four independent experiments (n = 4) ± SEM are shown, **p < 0.01, using ratio paired t-test. B A representative Western Blot showing levels of activated ERK (p-ERK1/2), ERK1/2 and as a loading control, Beta-actin. Protein lysates were from 48 h 3D acinar cultures derived from independent mice of the indicated genotype (WT, n = 3; Dusp5M^−/−, n = 2). C Representative images and quantification (D) of pancreatic acinar cells cultivated in a 3D in vitro model. (Scale bars, 100 µM.) Acinar cells were derived from either wild type (WT); n = 7 or Dusp5M^−/−; n = 6 mice and analysed morphologically based on the conversion of acinar cell clusters to ductal cyst structures that were comprised of a single layer of epithelial cells surrounding an empty luminal space. For each mouse and time point, the rate of transdifferentiation in four optical fields was determined. Cultures were either left untreated or stimulated by addition of epidermal growth factor (EGF; 25 ng/ml) to promote acinar cell transdifferentiation. Mean values are shown, ns not significant, **p < 0.01, ***p < 0.001, ****p < 0.0001, using unpaired t-test with Welch’s correction.