Figure 6. Loss of UBC-13 and CHN-1 affects protein levels of VPS-34. Schematic illustration of the IP experiment is shown at the left of panels (A, C, E, and G). Worm lysates were prepared from strains carrying flag-vps-34 and bec-1-myc (A, C), flag-vps-34, bec-1-myc, and myc-vps-15 (E) or flag-vps-34 and myc-vps-15 (G) generated by CRISPR-Cas9. VPS-34, BEC-1, and VPS-15 were detected by anti-FLAG and anti-MYC antibodies as indicated. Tubulin was the loading control for all blots. The graphs show relative protein levels, which were quantified using ImageJ (B) or ClinX Image Analysis (D, F, and H) as mean ± SD of 3 independent sets of experiments. Mutant datasets were compared with WT using the unpaired t test. **, P < 0.0001; all other points have P > 0.05.
Figure 6.

Loss of UBC-13 and CHN-1 affects protein levels of VPS-34. Schematic illustration of the IP experiment is shown at the left of panels (A, C, E, and G). Worm lysates were prepared from strains carrying flag-vps-34 and bec-1-myc (A, C), flag-vps-34, bec-1-myc, and myc-vps-15 (E) or flag-vps-34 and myc-vps-15 (G) generated by CRISPR-Cas9. VPS-34, BEC-1, and VPS-15 were detected by anti-FLAG and anti-MYC antibodies as indicated. Tubulin was the loading control for all blots. The graphs show relative protein levels, which were quantified using ImageJ (B) or ClinX Image Analysis (D, F, and H) as mean ± SD of 3 independent sets of experiments. Mutant datasets were compared with WT using the unpaired t test. **, P < 0.0001; all other points have P > 0.05.

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