Figure 2.

Analysis of TRC8 expression by Northern (A) and dot blot (B). (A) Gel resolved polyadenylated RNA (2 μg) from adult human tissues (CLONTECH) was hybridized under recommended conditions with a 1.5-kb 3′ TRC8 cDNA encompassing most of the TMs and the ring finger (bp 83–1623). A second, largely nonoverlapping probe (bp 1446–2212) yielded essentially the same pattern. The filter was exposed for 18 hr at −80°C. (B) A CLONTECH human RNA master dot blot was hybridized with the same probe as in A under recommended conditions and exposed for 15 hr. Final wash conditions were 0.1× standard saline citrate, 0.5% SDS at 55°C for 20 min. Signals were collected on a Molecular Dynamics PhosphorImager. Blank positions included B8, F5-F8, and G8. Central nervous system tissues (A1-A8 and B1-B7) included (in order) whole brain, amygdala, caudate nucleus, cerebellum, cerebral cortex, frontal lobe, hippocampus, medulla oblongata, occipital lobe, putamen, substantia nigra, temporal lobe, thalamus, sub-thalamic nucleus, and spinal cord. Musculature and digestive tissues (C1-C8) included heart, aorta, skeletal muscle, colon, bladder, uterus, prostate, and stomach. Secretory tissues (D1-D8) included testis, ovary, pancreas, pituitary, adrenal, thyroid, salivary, and mammary glands. Miscellaneous tissues (E1-E8 and F1-F4) included kidney, liver, small intestine, spleen, thymus, peripheral leukocytes, lymph node, bone marrow, appendix, lung, trachea, and placenta. Fetal tissues (G1-G7) included brain, heart, kidney, liver, spleen, thymus, and lung. All control spots (yeast and Escherichia coli RNAs, human Cot1, and total human DNAs) were blank (not shown).