Fig. 1

Antp and hTBP interact through polyglutamine stretches. Antp-hTBP interaction was determined using BiFC assays in transfected HEK293 cells. A Schematic representations of Antp and hTBP with their mutant versions. For Antp, the polyQ regions are indicated in shades of blue; YPWM, in purple; and the homeodomain (HD), in orange. For hTBP, the polyQ region is indicated in green. B BiFC assays revealed an interaction between Antp and hTBP (Venus reconstitution in green) and a reduction of BiFC between Antp and hTBP polyQ mutants. The number on the lower right corner indicates the percentage of cells showing the interaction. C Statistical analysis of BiFC-detected interactions between Antp-mutant versions and hTBP. AntpΔHD/^AAAA/Q5-hTBP showed no significant difference, in contrast, there is a highly significant difference (***) with AntpΔN/ΔPolyQ/Q9-hTBP. AntpQ6-hTBP interaction showed a significant difference (*). D Statistical analysis of BiFC-detected interactions between wild-type and mutant Antp-hTBP versions. hTBPΔQ40-Antp interaction showed a highly significant difference (***) in contrast with a non-significant difference for hTBPQ80-Antp. Antp or ΔN interactions with hTBP or its mutant versions have a highly significant difference in all pairs compared. pCAG-mCherry (red fluorescence) was used as an internal control. Three independent triplicates were analyzed using one-way ANOVA and the post-hoc test Tukey for mean comparison; all significant differences were obtained comparing to Antp-hTBP interaction. Significance is indicated as *** = p ≤ 0.001, ** = p ≤ 0.01, * = p ≤ 0.05, ns is not significant, and Error bars correspond to standard deviation. Scale bar, 30 µm