Widespread expression ofcnoin normal tissues and cells. (A) OriGene Technologies Northern blot showing expression of cno in mouse tissues. The blot was hybridized with a 738-bp probe amplified from gDNA that included the entire open reading frame of mouse cno (forward 5′-TAACGCAATCGCTCAGCACCGGAAG-3′, reverse 5′-ACAAGAGTCTCGCTGGAGTCCTCAT-3′). (B) By RT-PCR, cno is detected in RNA isolated from megakaryocytes and RNA from cultured C57BL/6J melanocytes. Total RNA was reverse transcribed using oligo-dT. The PCR primer sequences were as described in Figure 1B. (C) Quantitative real-time PCR analysis of wild-type and mutant tissues. Expression of cno in each tissue is presented relative to 18S RNA expression. The data are expressed as the ×±SD of 3 separate experiments. No significant differences in expression were seen in cno/cno tissues (□) versus +/+ tissues (▪). (D) Clontech human multiple tissue Northern blot hybridized with a human CNO probe (bp 396-917 of AK002092) amplified from fibroblast gDNA using forward primer 5′-GGCATGCTGGACATGCTTCGCGG-3′ and reverse primer 5′-CTATACATTCTACACTGAATCATG-3′. The origin of the high-molecular-weight band in skeletal muscle is unknown. The migration of RNA standards is shown to the left of panels A and D.