Densitometry of Hemoglobins of Heterozygous Thalassemic and Control Mice Analyzed by Immobilized Gradient Isoelectric Focusing
| Sample . | A.Percent Hemoglobin . | B.Relative Amount of Hemoglobin . |
|---|---|---|
| . | with Chain-1 . | with Chain-1 . |
| Wild type | 48.3 ± 1.2 | 100 |
| ζ−/+ | 20.5 ± 2.8 | 42.4 ± 5.8 |
| α1−/+ | 17.9 ± 1.2 | 37.1 ± 2.5 |
| Sample . | A.Percent Hemoglobin . | B.Relative Amount of Hemoglobin . |
|---|---|---|
| . | with Chain-1 . | with Chain-1 . |
| Wild type | 48.3 ± 1.2 | 100 |
| ζ−/+ | 20.5 ± 2.8 | 42.4 ± 5.8 |
| α1−/+ | 17.9 ± 1.2 | 37.1 ± 2.5 |
Samples from three individual mice of each genotype were scanned with a Shimadzu CS9000 densitometer and data analyzed using the Shimadzu CS Turbo software. All mice carry one normal Balb/c chromosome 11, which encodes α-globin chains-2 and -3. The mutated 129/SvEv chromosome encodes α-globin chain-1. The relative amount of hemoglobin with α-chain–1 (encoded by the mutant chromosome) is significantly reduced from normal in the insertional mutants, ζ−/+ and α1+/−. The data are presented (+/− SD). The difference between the mutants themselves is not significant (t-test). Thus, the presence of the inserted PGK-Neo cassette is accompanied by a reduction in expression of the α-gene in cis, both as a percent of the total hemoglobin present normally (column A) and relative to the normal product of the 129/SvEv chromosome (chain 1) (column B ). Expression of the α-genes encoded in trans are unaffected.