Differentiation of mouse intra-islet progenitor cells and immunoblotting of key transcription factors and islet markers under differentiation with activin, swertisin and presence of p-38 MAPK inhibitor.

<p>(A) depicts mIP cells cultured in complete media at day 0 which were then subjected to differentiation using activin-A and swertisin for 10 days. Bright field image shows, cells under differentiation on 8^th day at 20X magnification, and dithizone stained clusters on day 10^th. A fluorescent image represents immunostaining for insulin (green) and glucagon (red) in clusters from SFM/ITS, activin-a and swertisin groups. DAPI was used as nuclear stain. (B) shows immunoblotting of E-cadherin, N-cadherin, Ngn-3, P-p38, Native p-38 MAP kinase pathway proteins in presence and absence of MAP kinase pathway inhibitor SB203580. Ponceau S stain blot was shown as loading control. (C) shows immunoblotting of key differentiation pathway parameters that indicate the conversion of Mouse intra-islet progenitor cells into islet like clusters. Key transcription factors and MAP Kinase pathway proteins like P-p38, Erk1/2 Ngn-3, Pax-4, and Smad proteins under differentiation were monitored. Beta actin was used as loading control.