Figure 4.

Northern blot analysis of ACS4 mRNA in various rat tissues. Total RNA prepared from the indicated rat tissue was subjected to electrophoresis on a 1.5% agarose gel and blotted onto a nylon membrane. The blot was hybridized with a ³²P-labeled 2.2-kb SacI–XbaI fragment of pACS4 under stringent conditions. The filter was washed in 0.1× standard saline citrate (SSC) containing 0.1% SDS at 65°C for 30 min and exposed to Kodak XAR-5 film with an intensifying screen at −80°C for 48 h. The lanes were loaded with 15 μg of total RNA isolated from liver (li), brain (br), lung (lu), heart (He), adipose tissue (At), small intestine (Si), skeletal muscle (Sk), spleen (Sp), kidney (Ki), adrenal gland (Ad), testis (Te), epididymis (Ep), seminal vesicle (Se), ovary (Ov), and uterus (Ut). The same samples in A were subsequently hybridized with a control probe for rat glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (CLONTECH) and exposed to Kodak XAR-5 film with an intensifying screen at −80°C for 20 h. The autoradiograph shown is a representative of five experiments that gave essentially identical results.