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Fig. 6. Suppression of hypoxia-induced cell death by over-expression of
CaHO-1. (A) Confirmation of the stably transfected CAB cells by
semi-quantitative RT-PCR using T7 primer and CaHO-1-specific primer
HO1-R4. A band (0.67 kb) was only detected in CAB cells stably transfected
with plasmid pcDNA3.1 carrying the full-length ORF of HO-1 gene;
β-actin was employed as an internal control. Lane M, DL2000 DNA marker;
lane 1, CAB/pcDNA3.1 cell; lane 2, CAB/pcDNA3.1-HO-1 cell. (B) Over-expression
of HO-1 in stably transfected CAB cells under hypoxic treatment.
Total RNA was isolated from CAB/pcDNA3.1 and CAB/pcDNA3.1-HO-1 cells after 4
days of hypoxia (1% O2). The transcription of CaHO-1 was
then evaluated by semi-quantitative RT-PCR (lane 2). CAB/pcDNA3.1 cells were
employed as a control (lane 1). (C) Morphological observation of
hypoxia-induced cell death under phase contrast microscope. (D) Viability of
CAB/pcDNA3.1 and CAB/pcDNA3.1-HO-1 cells under the hypoxia–reoxygenation
treatment. Cell viability was examined by a modified MTT assay using a CCK-8
kit each day (see Materials and methods). The experiments were repeated at
least three times and the symbols represent the mean values of triplicate
wells, with standard deviations.
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