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From: Lentiviral Vector-Mediated PAX6 Overexpression Promotes Growth and Inhibits Apoptosis of Human Retinoblastoma Cells Invest. Ophthalmol. Vis. Sci ;52(11): doi: /iovs Figure Legend: Q-PCR analysis (a, b) and Western blot analysis (c) for the assessment of the PAX6 overexpression in two human retinoblastoma cell lines, Y79 and SO-Rb50, after transfection by lentiviral vectors. (a, b) Five days after transfection, total RNA was extracted from the cells of the study groups (transfected with PAX6-GFP lentiviral vectors) (pGCL-GFP-PAX6), cells of the negative GFP control groups (transfected with GFP lentiviral vectors) (pGCL-GFP), and cells of the negative control groups without transfection (Mock). Q-PCR was performed to measure the relative PAX6 mRNA levels. The amount of each product was normalized to human β-actin mRNA levels. Each value represented the mean ± SD of three replicates. PAX6 mRNA levels were significantly higher in the study groups (**P < 0.001). (c) Seven days after transfection, protein expression levels of PAX6 were assessed by Western blot analysis using the anti-human PAX6 antibody, applying the enhanced chemiluminescence method and quantification with the analysis software. The levels of the PAX6 protein were significantly higher in the study groups (pGCL-GFP-PAX6) than in the control groups (pGCL-GFP and Mock). In comparison, the GAPDH protein did not vary markedly among the groups. (Note: the protein band at 46 kDa represents the PAX6 protein, and the protein band at 74 kDa represents the PAX6-GFP fusion protein.) Date of download: 10/18/2017 The Association for Research in Vision and Ophthalmology Copyright © All rights reserved.
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