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. 2000 May 2;97(10):5562–5567. doi: 10.1073/pnas.100118597

Figure 2.

Figure 2

β4 mRNA is abundant in brain. (a) PolyA+ RNA Dot blot (CLONTECH) of 50 human tissues were hybridized at high stringency with a 32P-labeled β4 (681 bp, AccI–AccI fragment) probe. Tissues are: whole brain (1A), amygdala (2A), caudate nucleus (3A), cerebellum (4A), cerebral cortex (5A), frontal lobe (6A), hippocampus (7A), medulla oblongata (8A), occipital lobe (1B), putamen (2B), substantia nigra (3B), temporal lobe (4B), thalamus (5B), nucleus accumbens (6B), spinal cord (7B), heart (1C), aorta (2C), skeletal muscle (3C), colon (4C), bladder (5C), uterus (6C), prostate (7C), stomach (8C), testis (1D), ovary (2D), pancreas (3D), pituitary gland (4D), adrenal gland (5D), thyroid gland (6D), salivary gland (7D), mammary gland (8D), kidney (1E), liver (2E), small intestine (3E), spleen (4E), thymus (5E), peripheral leukocyte (6E), lymph node (7E), bone marrow (8E), appendix (1F), lung (2F), trachea (3F), placenta (4F), no sample (5F-8F, G8), fetal brain (G1), fetal heart (G2), fetal kidney (G3), fetal liver (G4), fetal spleen (G5), fetal thymus (G6), fetal lung (G7), 100 ng yeast total RNA (H1), 100 ng of yeast tRNA (H2), 100 ng of E. coli rRNA (H3), 100 ng of E. coli DNA (H4), 100 ng of poly r(A) (H5), 100 ng of human Cot 1 DNA (H6), 100 ng of human DNA (H7), and 500 ng of human DNA (H8). The amount of RNA in each dot is between 89 and 514 ng, due to a normalization with respect to ubiquitously expressed mRNAs. (Inset) Internal dot blot standard of β1, β2, and β4 transcripts. The internal standard shows a detection limit of 0.01 fmol or 6 × 106 transcripts. (b) Quantification of Dot blot signals using phosphorimaging. The signals were background subtracted and normalized to equal amounts of RNA. Absolute transcript numbers were obtained using the signals from the internal standard assuming similar hybridization efficiencies. (c) Northern Blots were hybridized and visualized as in a. The signals on the blot with fetal tissues are stronger than for the blot with adult tissues (c); however, for a comparison of expression levels, compare signals on the same blot (a and b). (d) β4 gene is localized in chromosome 12q15–21. Double labeling of chromosome 12 with a chromosome 12 centromere-specific probe (red signal) and genomic β4 probe (green signal) (n = 10 measurements).