Identification of differentially expressed genes after partial rat liver ischemia/reperfusion by suppression subtractive hybridization

Figure 3 Second screening of SSH-clones and ‘Atlas Arrays’. Two identical sets of Southern blots and two Atlas-Arrays were hybridized with complex probes of four pooled C-lobes or matched I-lobes after partial WI/R (1 h/2 h). A: Typical results of two corresponding Southern blots. The upper membranes were screened with the C-lobe probe (pool-C) and the lower membranes show the result after hybridization with the I-lobe probe (pool-I) pool-C/pool-I ratios of signal intensities after densitometry: 0.4 (1, 4), 1.1 (2, 3), 0.2 (5), 2.0 (6), 2.6 (7), 2.8 (8), 2.2 (9), 7.0 (10). β-Actin-positive controls (150 ng/lane, 2200 ng/lane); B: Ethidium bromide-stained agarose gels before blotting; C: Distribution of pool-C/pool-I ratios of all 89 analyzed genes; D: Exemplary hybridization results of the two cDNA arrays after 4 h exposition. Numbered black or white arrows indicate spots of up- or down-regulated genes in the I-lobe cDNA-pool, which were among others chosen for Northern blot hybridizations. Jun (1), Hsp27 (2), Hsp70 (3), IL-1RI (4) and Fgb (5). Arrow heads point to spotted housekeeping gene cDNAs; β-Actin (6), ribosomal protein S29 (7), 1 spotted genomic DNA to assess contaminations in the RNA preparations, used for probe synthesis.