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Volumn 277, Issue 5323, 1997, Pages 242-245

Pericyte loss and microaneurysm formation in PDGF-B-deficient mice

Author keywords

[No Author keywords available]

Indexed keywords

PLATELET DERIVED GROWTH FACTOR B;

EID: 0030756325     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.277.5323.242     Document Type: Article
Times cited : (1852)

References (38)
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    • lcz heterozygote embryos being either PDGF-B -/-or PDGF-B +/+ were identified by PCR.
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    • 4, pH 7.5. Heated probes (see below) were added at a concentration of 3 to 8 μg/ml in hybridization solution, and the sections were incubated overnight. Washes after hybridization were carried out in 1 × SSC, 50 to 55% formamide, and 0.1% Tween 20. The entire process was performed at 65° to 72.5°C (the higher stringency was used for the PDGF-B probe). Production of digoxygenin (DIG)-labeled RNA probes and detection of the probes on sections with an alkaline phosphatase-conjugated antibody were accomplished with DIG-labeled uridine triphosphate, the DIG RNA Labeling Kit, and the DIG Nucleic Acid Detection Kit (Boehringer Mannheim) according to the manufacturer's instructions. PDGF-B sense and antisense probes were generated from a 0.8-kbp mouse cDNA containing the full-length coding sequence cloned in pBS-SK. PDGF-Rβ probes were generated from a 461-bp Sac I fragment cloned in pGEM-2. Figure 2 shows the use of antisense probes on 14-μm thick sections. Photography was done with a Nikon photomicroscope. Unstained sections and Normaski optics were used to allow for good sensitivity and resolution. Controls for the specificity of the in situ hybridization signals included sense probe hybridizations and PDGF-B antisense probe hybridization to PDGF-B -/-embryos. No hybridization signal was obtained in any of these controls (14). PDGF-Rβ expression was studied on 206 sections taken from seven littermate pairs of PDGF-B +/+ and -/-embryos. PDGF-B expression was studied on 108 sections taken from two PDGF-B +/+ and one PDGF-B -/-embryos. Representative examples of these analyses are shown in Figs. 2 and 3.
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    • note
    • lcz mice; M. Mercola, J. Escobedo, P. Soriano, and W. D. Richardson for probes; and the Swedish Cancer Foundation, the Swedish Medical Research Council (MFR), the IngaBritt and Arne Lundberg Foundation, and the Göteborg University for financial support.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.