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. Author manuscript; available in PMC: 2008 Nov 26.
Published in final edited form as: Mol Endocrinol. 2006 Apr 6;20(8):1796–1809. doi: 10.1210/me.2005-0327

Fig. 3. Analyses of the Recombination Frequency of the T/ebp-Floxed Allele, T/ebp Expression Levels, and Serum TSH Levels.

Fig. 3

A, Schematic diagram of the location of primers P1, P2, and P3 (arrows) that were used for determination of recombination frequency, and primers a and b (shaded arrowheads) that were used for quantitation of T/ebp mRNA levels by real-time PCR. The thick line indicates the sequence in between two XhoI sites that is present only in the floxed allele derived from the targeting vector and differs from that of KO allele. The P2/P3 primer pair generates an amplicon (164 bp) only from the unrecombined floxed allele, but not from the recombined deleted allele because of the loss of the P2 primer binding sequence following Cre/loxP excision of exon2, nor the KO allele because P2 primer sequence does not exist. The P1/P3 primer pair generates an amplicon (165 bp) from the recombined deleted allele, but not from the unrecombined floxed or KO allele because of the large amplicon size. The diagram is not drawn to scale. Restriction enzyme sites: Xh, XhoI. B, Frequency of T/ebp-floxed gene recombination in T/ebp(fl/fl); TPO-Cre mice. The ratio of the recombined allele is shown as a percentage of the total floxed (unrecombined and recombined together) alleles, which was set at 100%. Values are the means ± SE. C, Expression levels of T/ebp mRNA in thyroid glands. All mRNA levels are normalized to 18S rRNA and presented as a relative ratio to the level in T/ebp(fl/fl) mice in each age group set to 1.0. Values are the means ± SE. D, Serum TSH levels in T/ebp(fl/fl), T/ebp(fl/fl); TPO-Cre, and T/ebp(fl/ko) mice. Each dot represents a TSH level for single mouse, and horizontal bars represent the mean values. Asterisks indicate mice that demonstrated histological atrophic/degenerative abnormalities in their thyroids as described in the text. NS, Statistically not significant; mo, month.