Fig. 6

LTA4H was upregulated by ACR and induced vascular alteration in akr7a3^−/− zebrafish. (A–B) Representative confocal images and quantification of hyaloid blood vessel diameters in larvae at 5dpf. The increased diameter of hyaloid vasculature in akr7a3^−/− zebrafish was significantly reduced after LTA4Hi incubation. White scale bar = 30 μm, n = 18–20. (C–D) RT-qPCR analysis of LTA4H expression. LTA4H was significantly increased in akr7a3^−/− zebrafish larvae. Incubation with CAR reduced the expression of LTA4H non-significantly. After incubating akr7a3^+/+ zebrafish larvae with ACR, an increase in LTA4H expression was observed. However, co-incubation with ACR and CAR was able to inhibit the elevated expression of LTA4H. n = 6 clutches with 30 larvae. (E–G) RT-qPCR analysis data revealed an increased expression of AhR (E), while Junb-a (F) and Junb-b (G) exhibited decreased expression. n = 6 clutches with 30 larvae. (H–I) Docking analysis showed ACR (I) shared same binding pocket with Indirubin (H). ACR formed a hydrogen bond with the amino residue SER336 of AhR. Dash line = hydrogen bonds. For statistical analysis one-way ANOVA or Student's t-test was applied. The bars indicate values of mean ± SD. AhR, Aryl hydrocarbon receptor; Junb-a, JunB Proto-Oncogene a; Junb-b, JunB Proto-Oncogene b.