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Fig. 2 | Stem Cell Research & Therapy

Fig. 2

From: Puromycin-based purification of cells with high expression of the cytochrome P450 CYP3A4 gene from a patient with drug-induced liver injury (DILI)

Fig. 2

Selection of puromycin-resistant cells. A Scheme for selection of puromycin-resistant cells (green) from a mixed population of immortalized cells. For selection, cells were treated with puromycin (3 μg/ml) for 3 days. B, C Phase-contrast micrograph of puromycin-selected cells. B: Low-power view. C: High-power view. DH Quantitative RT-PCR analysis of the genes for AFP (D), ALB (E), CYP1A2 (F), CYP2B6 (G), and CYP3A4 (H) after the puromycin treatment. The immortalized differentiated cells (Fig. 1A) were exposed to puromycin at a final concentration of 3 μg/ml for 3 days. RNAs were isolated from the cells 1 week after the puromycin treatment. (−): no puromycin treatment, (+): puromycin treatment. The amounts of each gene at the cells without puromycin treatment were regarded as equal to 1. Statistical analysis was performed using the unpaired two-tailed Student's t-test. **P < 0.01. IR Quantitative RT-PCR analysis of the puromycin-selected immortalized cells after propagation (Puro-cell), HepG2 cells, and hepatocytes (Liver). I: AFP, J: ALB, K: CYP1A2, L: CYP2B6, M: CYP3A4, N: CPS1, O: OTC, P: OATP1B1, Q: OATP1B3, R: HNF4A. S Immunocytochemistry of puromycin-selected cells with the antibodies to ALB (green) and AFP (red). T Immunocytochemistry of puromycin-selected cells with the antibodies to CYP3A4 (green). U CYP3A4 enzymatic activity in HepG2 cells, Caco2 cells, and the puromycin-selected cells

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