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Fig. 6 - Supplemental 1 Stable foxp1a and foxp1b mutant alleles lead to reduced transcript and protein expression. (a) qRT-PCR showing significantly reduced foxp1a mRNA expression in homozygous foxp1aed116 mutants compared to heterozygotes and wild-type siblings. (b) Expression of other foxp1 family genes in foxp1aed116 mutants shows no evidence of compensatory upregulation. (c) Western blot of Foxp1 protein in foxp1a wild-type (+/+) and foxp1aed116 mutant caudal fin lysates. Foxp1 protein is markedly reduced in mutants. β-Actin serves as a loading control. (d) qRT-PCR showing significantly reduced foxp1b mRNA in foxp1bed125 mutants compared to heterozygotes and wild-type siblings. (e) Expression of other foxp family genes (foxp1a, foxp2, foxp3a, foxp3b, foxp4) in foxp1bed125 mutants. foxp3a and foxp3b are significantly downregulated in heterozygotes; however, this likely reflects defective amplification in those samples, as homozygous mutants show no change. (f) Western blot of Foxp1 protein in foxp1b wild-type (+/+) and foxp1bed125 caudal fin lysates, showing reduced Foxp1 protein in mutants. β-Actin was used as a loading control. Error bars represent standard error of the mean (SEM). ***p<0.001, by unpaired two-tailed t-test.