FIG. 4.

The cells originating from the reprogramming protocol consist of at least two populations, one that is enriched for glucagon and the other enriched for insulin. (a) Human primary dermal fibroblasts were stably infected with a lentivirus containing green fluorescent protein (GFP) under the control of the glucagon gene promoter. Left panel—control hDFs were infected with LacZ adenovirus, not Pdx1, and were not exposed to Romi and 5-AzC=. Right panel—cells undergoing the reprogramming protocol. Cells were sorted by FACS into cells expressing GFP and GFP-negative cells. (b) Real-time PCR analysis of glucagon and insulin in hDFs undergoing the protocol. Data are expressed relative to GAPDH mRNA. Values are the mean±S.E.M from three separate experiments, each analyzed in duplicate. *, statistical significance with P<0.05 value using a Student's t-test. (c) Fluorescent in situ hybridization with insulin (green) and glucagon (red) probes in cells under the reprogramming protocol.