Figure 3 | Laboratory Investigation

Figure 3

From: Reprogramming liver-stem WB cells into functional insulin-producing cells by persistent expression of Pdx1- and Pdx1-VP16 mediated by lentiviral vectors

Figure 3

Profiles of short-term gene expression of Pdx1 or Pdx1-VP16. Total RNA was extracted by trizol reagent and treated with DNAse. A total RNA (2 μg) was reversely transcribed and PCR was performed at 35 cycles and 1/5 of PCR products were analyzed by 1.8% of agarose gel and visualized with ethidium bromine. All primers were designed to cross intron (s) except for MafA and ngn3 (genes have only one exon) and the primer sequences are provided in Table 1. Rat insulinoma INS-1 (823/13) cells serve as positive controls for beta-cell gene expression. Parental WB cells transduced with LV-GFP serve as baseline for lentiviral vector control. Total RNA samples without RT were performed to exclude DNA contamination. Actin serves as house-keeping gene control to give relative expression levels in each sample. Pdx1 and Pdx1-VP16 plasmid were used as positive controls.

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