Journal: Scientific Reports
Article Title: An inhibitor of fibroblast growth factor receptor-1 (FGFR1) promotes late-stage terminal differentiation from NGN3+ pancreatic endocrine progenitors
Figure Lengend Snippet: C8 augments important maturation markers specifically in INS-producing β cells. Quantitative RT-PCR analysis was performed to measure the relative expression levels of the β cell lineage markers INS , UCN3 , and GCK and the transcription factors PDX1 , NKX6.1 , and MAFA from sorted Venus+ and NC cells with or without treatment with C8 and/or bFGF. Briefly, #9–15 hiPSCs were differentiated as described above and treated with 5 μM C8 or 5 μM C8 and 50 ng/mL bFGF in the third stage of differentiation. After differentiation, samples were dissociated with TrypLE and sorted with an SH800Z cell sorter (Sony) for Venus+ and NC cells. Fifty cells from each group were sorted into 5 μL FCP reagent, which is a lysate reagent of the QIAGEN Fast Lane cDNA kit. For cDNA synthesis, 2 μL of the samples were used, with 0.5 μL of the 10 μL cDNA samples used for quantitative RT-PCR. Error bars indicate SD, n = 3. *p
Article Snippet: The number of Venus- and mCherry-expressing cells was analysed in an SH800Z cell sorter (Sony, Tokyo, Japan).
Techniques: Quantitative RT-PCR, Expressing