Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2011 Nov 14;195(4):689–701. doi: 10.1083/jcb.201107029

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2011 Kurihara et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

PMC Copyright notice

Figure 4. — HIF-1α, but not HIF-2α, regulates MIF in astrocytes. (A) Mif mRNA expression was monitored by real-time RT-PCR and plotted on a scatter plot at P3, 6, 9, 11, 14, 21, and 28 early postnatal stages of hyaloidal vessel regression in isolated retinas. Note that significant down-regulation is observed at P14 compared with P3 (n = 8). (B) Immunohistochemistry with MIF and GFAP antibodies in P14 retinal ganglion cell layer is shown. Note that MIF expression is colocalized with astrocytes in the ganglion cell layer. (C) Mif and F4/80 mRNA expression in Vhl/Hif-2α double mutants at P14. Note that the ectopic stabilization of HIF-1α in retinal astrocytes results in simultaneous (and significant) up-regulation of Mif and F4/80 (n = 4). KO, knockout. (D) Immunocytochemistry for MIF was performed on cultured C8-D1A murine astrocyte cells. Note that MIF is robustly expressed in the cytoplasm (visualized with phalloidin labeling) of C8-D1A cells (all cells are counterstained with DAPI). (E) Relative mRNA expression of Hif-1α, Hif-2α, Vegf, and Mif in C8-D1A cells transfected with CA–Hif-1α, CA–Hif-2α, or Gfp. Note that the transfection of CA–Hif-1α induces Hif-1α in C8-D1A (top left), and CA–Hif-2α induces robust Hif-2α expression (top right). Vegf is up-regulated significantly (4.9-fold; P = 0.00024) when C8-D1A cells are transfected with CA–Hif-2α. Mif is significantly (1.9-fold; P = 0.000057) up-regulated by CA–Hif-1α (n = 11–12). (F) VEGF and MIF-secreted protein levels were measured from supernatants of CA–Hif-1α or CA–Hif-2α–transfected C8-D1A cells using quantitative sandwich ELISAs (n = 5–6). VEGF secretion is significantly (2.7-fold; P = 0.009) up-regulated by CA–Hif-2α (top). MIF is significantly (7.6-fold; P = 0.03) up-regulated by CA–Hif-1α (bottom). (G–J) Representative hyaloidal vasculature (G and I) and number of branching points (H and J) in HIF-1α (G and H) or HIF-2α (I and J) mutants are shown at P9 (n = 6). Note that in HIF-1α mutants, delayed hyaloidal regression compared with control littermate is observed in early postnatal stages. n.s., not significant. *, P < 0.05; **, P < 0.01. Error bars indicate mean ± SD. Bars: (B) 20 µm; (D) 50 µm; (G and I) 2,000 µm.