Figure 3
From: Definition of a Skp2-c-Myc Pathway to Expand Human Beta-cells
P27KIP1 decreases the proliferative response of human β-cells.
(A) Representative immunoblot of human islets transduced with a control adenovirus (CTL) or with Ad.cdk6 and cyclin D3 (C6 + D3), or with Ad.cdk6 and cyclin D3 with shRNA to silence p27kip1 (C6 + D3 + shp27kip1), or with shRNA to silence p27kip1 (shp27kip1). (B) Densitometric analysis of p27kip1 immunoblots. Experiments were repeated five times. *p < 0.05. (C) Dispersed human islets from normal donors were transduced with control adenovirus (“CTL”) or with Ad.cdk6 and Ad.cyclin D3 (“C6 + D3”) or with Ad.cdk6 and cyclin D3 with shRNA to silence p27kip1 (C6 + D3 + shp27kip1). Immunolabelling for Ki67 is shown in red, insulin in white and DAPI in blue. (D) Quantification of data in C at 72 hours after transduction. Bars indicate mean ± SEM of the % of the proliferation with cdk6 and cyclin D3 (n = 5). (E) Dispersed human islets from normal donors were transduced with control adenovirus (“CTL”) or with Ad.cdk6 and cyclin D3 with shRNA to silence p27kip1 (C6 + D3 + shp27kip1). Immunolabelling for γ-phospho-H2AX (H2AX) is shown in red, insulin in white and DAPI in blue. (F) Quantification of H2AX immunolabelling at 72 hours after transduction (n = 5). Bars indicate mean ± SEM of the % of H2AX positive β-cells.
