Figure 3. Autoradiographs of a Northern blot showing the effect of anesthesia and surgery on hepatic heat-shock protein-72 (hsp72), inter-alpha-trypsin inhibitor (alpha-Ti), and tubulin steady-state RNA levels. Total RNA (20 micro gram/lane) was isolated from serial pig liver biopsy samples at baseline (time 0), and at 1, 2, 3, 4, and 6 h after baseline. The RNA was electrophoretically resolved, blotted, probed with hsp72 cDNA, and autoradiographed. The same blot was subsequently stripped and reprobed twice to determine alpha-Ti and tubulin RNA levels. The positions of the 18 S and 28 S ribosomal RNA subunits derived from visual inspection of the ethidium-bromide-stained gel are indicated. Analysis of the tubulin RNA level was included to confirm the lane-to-lane uniformity of RNA loading and was used as a standard to allow formal normalization of the hsp72 and alpha-Ti signals. RNA processed for this Northern blot and the hepatic nuclei used for the runoff analysis shown in Figure 1were obtained from the same animal.