Figure 3

Cell cycle reentry in primary human islets following knockdown of RB and p53 pathway members. Primary human islets were electroporated with siRNAs against non-targeting control, or combinations of RB, p107 (L-003298), p130 (L-003299), p21 (L-003471), p53 (L-003329) and ARF/CDKN2A (L-011007). Islets were cultured in the presence of 5-ethynyl-2′-deoxyuridine (EdU) for 7 days following electroporation then fixed and stained with DAPI and for EdU. (A) Images showing EdU incorporation in Islets with nuclei stained with DAPI (blue, left), EdU (red, middle) and a composite image (right). (B) Graph showing percentage of EdU incorporating cells in islets following transfection. Total cells counted in the representative experiment presented are: siControl (245); Rb + p53 (535); Rbfamily + p53 + p21 (456); Rbfamily + p53 + p21 + Arf (834). The experiment was repeated 3 separate times.