Proindy

At 5 dpf, zebrafish larvae were incubated in treatment solutions for 3 h prior to imaging, and for 3 h during imaging, using 96-well ProxiPlates (PerkinElmer, 6006290). Zebrafish larvae were treated with 190 FDA-approved compounds using a Tocris small-molecule library (Tocris Bioscience, Cat. No. 7200). The library contained 10 mM stocks dissolved in dimethyl sulfoxide (DMSO), which we diluted 1000 × in egg water to a 10 µM final concentration. The imaging experiments also included untreated larvae in egg water and larvae treated with 1 µl/ml DMSO as a vehicle control. The effects of FDA-approved drugs were compared to previously obtained results^{26 (link)}, with 10 µM cyclosporine A (CsA, Enzo Life Sciences), 1 µM tacrolimus (FK506, Enzo Life Sciences), 1 µM rapamycin (Santa Cruz Biotechnology) and 5 and 10 µM proINDY (Tocris Bioscience).

Using 96-well ProxiPlates (PerkinElmer, 6006290), 5 dpf zebrafish larvae were incubated in treatment solutions for 3 h before imaging, and for 3 h during the imaging session. Zebrafish larvae were treated with 190 FDA-approved compounds using a Tocris small-molecule library (Tocris Bioscience, Cat. No. 7200). The library contained 10 mM stocks dissolved in dimethyl sulfoxide (DMSO), which was diluted 1000 × in egg water to a 10 µM final concentration. The control groups consisted of untreated larvae in egg water and larvae treated with 1 µl/ml DMSO serving as vehicle control. The behavioral effects of FDA-approved drugs were compared to previously obtained results^{20 (link)} with 10 µM cyclosporine A (CsA, Enzo Life Sciences), 1 µM tacrolimus (FK506, Enzo Life Sciences), 1 µM rapamycin (Santa Cruz Biotechnology), 5 and 10 µM proINDY (Tocris Bioscience) and a rescue experimental group typically consisting of calcineurin inhibitor along with proINDY. The rescue group for the current study consisted of four combinations—10 µM CsA + 5 μM proINDY, 10 µM CsA + 10 μM proINDY, 1 µM tacrolimus + 5 μM proINDY and 1 µM tacrolimus + 10 μM proINDY.