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. 2020 Sep 4;142(39):16825–16841. doi: 10.1021/jacs.0c07726

Figure 6.

Figure 6

Probing UCHL1 activity with 8RK59 in morpholino (MO)-mediated UCHL1 knockdown zebrafish embryos. (A) Schematic overview of labeling UCHL1 activity with 8RK59 in zebrafish embryos with/without UCHL1MO injection. (B) Bright field images of 2 dpf zebrafish embryos without injection (uninjected) after injection with standard control morpholino (control MO) or with two independent UCHL1MOs (UCHL1MO1 or UCHL1MO2). (C) Quantification of the curly tail phenotype in 2 dpf zebrafish embryos with/without UCHL1MO injection. (D) WB of UCHL1 in 2 dpf zebrafish embryos with/without UCHL1MO injections. WB for tubulin was included as a loading control. The expression levels of UCHL1 normalized to tubulin are indicated below. (E) Probing UCHL1 activity with 5 μM 8RK59 in 4 dpf zebrafish embryos with/without UCHL1MO injections. (F) Statistical analysis of 8RK59 signal in 4 dpf zebrafish embryos with/without UCHL1MO injections. The intensity of 8RK59 was measured in three zebrafish embryos of each group by calculating the pixel sum in the GFP channel of both the head and tail areas. *, P < 0.05, **, P < 0.01, ***, P < 0.001, and two-way ANOVA.