Figure 5.

EIF4A2 variants differently modulate dpp-induced eye phenotypes and failed to rescue the fly eIF4A LOF lethality

(A) Overexpression of dpp in the fruit fly eye using GMR-GAL4. (i) Empty control eye shows smooth eye surface and (v) hexagonal arrangement of ommatidia in the imprint. (ii) Expression of dpp results in roughened eye surface and (vi) disorganized ommatidial arrangement. (iii) Expression of fly eIF4A in the background of GMR-GAL4>dpp shows slight reduction in roughened eye surface (iv) and better ommatidial arrangement. (v) Expression of EIF4A2 WT in the background of GMR-GAL4>dpp did not make any changes to the roughened eye surface (iv) and ommatidial arrangement.

(B) Expression of EIF4A2 variants in the background of GMR-GAL4>dpp. (i) Expression of EIF4A2 p.Gly364Glu did not make significant changes to the roughened eye surface and (v) disorganized ommatidial arrangement in the imprint. (ii and iii) Expression of EIF4A2 p.Leu344Phe and EIF4A2 p.Thr243Ile show roughened eye surface (vi and vii) and exacerbates the disorganized ommatidial arrangement. (iv) Expression of EIF4A2 p.Thr216Ile did not made any changes to the roughened eye surface (viii) and ommatidial arrangement.

(C) Quantification of misshaped ommatidia. (i and ii) Expression of EIF4A2 p.Leu344Phe and EIF4A2 p.Thr243Ile shows significant increase in the total number of misshaped ommatidia and percentage of misshaped ommatidia per field of view compared to the GMR-GAL4>dpp and GMR-GAL4>dpp; EIF4A2 WT controls. One-way ANOVA followed by Tukey’s post hoc test was performed for the statistical analysis. Data shown mean ± SEM with sample size of total number of male and female flies shown in figure. Significance shown as ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001.

(D) Knockdown of eIF4A in the GMR domain results in pupal lethality. Expression of human EIF4A2 WT in this background results in complete rescue of pupal lethality, whereas the variants fail to rescue the lethality.