Hs

mhsp70i (A) rhsp70i mhsp70c

mhsp70i (A)

rhsp70i mhsp70i (B)

mhsp70c

Fig. 6. (A) Western blot analysis. Proteins from hearts of transgene-positive and transgene-negative mice were probed with polyclonal and monoclonal antibodies. The polyclonal antibody recognizes constitutive (HSP 70c) and inducible (HSP 70i) forms of the 70-kDa heat shock protein. The monoclonal antibody recognizes only HSP 70i. A strong HSP 70i signal is seen in the hearts of transgene-positive mice but not in transgene-negative mice. (B) Northern blot analysis. RNA isolated from heart and skeletal muscle of transgene-positive (lane 1) and transgene-negative (lane 2) mice was analyzed by hybridization of 32P-labeled rHSP 70i cDNA. High levels of rHSP 70i were seen in heart and skeletal muscle of transgene-positive animals but not in transgene-negative animals. In lane 3, samples from transgene-negative mice subjected to whole body heat stress (8 hr after 42° for 15 min) were loaded for comparison. Heat stress has a greater effect in inducing mHSP 70i in skeletal muscle than in heart. Both forms (A and B) of mHSP 70i are indicated and the size of the chimeric transgene is different from that of the endogenous mouse HSP 70i transcript.

buffer [composed of (in mW): 118 NaCl, 24 NaHC03, 4.7 KC1, 1.2 KH2P04, 1.2 MgS04, 1.7 CaCl2, 10 glucose, and gassed with 95% 02:5% C02, pH 7.4 at 37°] by the retrograde Langendorff method at a constant perfusion pressure of 80 cmt^O6'12 (Fig. 7A). A small incision is made at the main trunk of the pulmonary artery to drain coronary effluent. The effluent is collected to measure the

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