Fig. 4

Fig. 4 P62 protein levels are increased in Dcc-enriched fractions of ids mutant fish-derived protein lysates.

A Graphical representation of the workflow adopted for the fractional precipitation assay (created with Biorender). Briefly, 2 dpf dissected heads were homogenized and subjected to first centrifugation; then, the supernatant was loaded on top of a sucrose column to be submitted to ultracentrifugation. Finally, fractions were retrieved from the top (low density) to the bottom (high density) of the sucrose gradient. B Representative western blot depicting wild-type and ids mutant fish lysates subjected to fractional precipitations. Fractions are reported from 1 to 7, with bold numbers highlighting the fraction with higher Dcc enrichment (fraction 3 for control and fraction 1 for ids mutant-derived lysates). The distributions of P62, N-cadherin (Cdh2), Gapdh (glyceraldehyde-3-phosphate dehydrogenase), Rab7 (Ras-associated binding protein 7), and Lc3-II were analysed. While the majority of markers were distributed in the same way in the two genotypes, P62 was particularly enriched in the same fraction displaying high Dcc protein levels only in ids mutant conditions.