QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

Help viewing high resolution images Return to article

Fig. 2. CoCl₂ induced HIF-1 and its target gene expression in MSCs. (A) HIF-1 immunostaining observed in MSCs with confocal microscopy after 3 and 72 hours of 100 µM CoCl₂ treatment. In the third image from the left, nuclei were counterstained with Propidium Iodide (PI, in red). Bars, 20 µm. (B) HIF-1 expression analysed by western blotting of cytoplasmic and nuclear extracts from MSCs after 3 hours of CoCl₂ treatment. (C) Expression of HIF-1 target genes analysed by real-time RT-PCR after 6 and 24 hours of CoCl₂ treatment, n=3, *P<0.05, univariate Student's t-test. (D) Western blot of EPO and VEGF expression after 72 hours of CoCl₂ treatment. Actin expression was used as protein loading control. (E) MSC morphology observed after 72 hours of 100 µM CoCl₂ treatment with or without 1.25 nM echinomycin (objective 20x). (F) Luciferase activity measurement after 36 hours of 100 µM CoCl₂ ± 1.25 nM echinomycin treatment. Firefly luciferase activities were normalized with the internal control Renilla luciferase activity. The fold induction represents the luciferase activity observed in transfection with the HRE-pGL3SV40 plasmid relative to the empty plasmid (pGL3SV40). Values are represented as fold induction compared with control ± s.e.m., n=3, *P<0.05, PLSD Fisher.

Return to article