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Fig. 1.   Detection and measurement of myosin heavy chain (MHC) IIx mRNA in human muscle. Presented is an RNase protection assay from one subject with ³²P-labeled riboprobes, yielding a 122-base pair protected fragment for human MHC IIx mRNA and a 316-base pair fragment for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA. Total RNA (2 µg) from each individual sample was incubated with MHC IIx and GAPDH riboprobes, digested with RNase, resolved on polyacrylamide gels, and identified by phosphorimager. Lane A: before training (day 1); lane B, 3 h after a single training bout (day 1 + 3 h); lane C, after the 7th training bout (day 7 + 3 h); lane D, after 6 days of training (day 7). Individuals with high initial levels of the transcript, such as the subject presented in this figure, demonstrated a trend for MHC IIx mRNA to decrease at day 7 (lane D) and to demonstrate relatively large decreases at day 7 + 3 h (lane C; see Fig. 4). There was, however, no mean reduction in MHC IIx mRNA at day 7 and a mean reduction of 30% (P < 0.05) 3 h after the final training bout (day 7 + 3 h) in the 9 subjects studied.