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. 2018 May 17;115(23):E5317–E5325. doi: 10.1073/pnas.1804091115

Fig. 2.

Fig. 2.

Wnt signaling increases endocytosis, and arginine-methylated proteins are sequestered by microautophagy. (A and A′) meArg colocalized with LysoTracker. Panel A′ was treated with Wnt3a for 15 min. (B) Wnt increased colocalization of meArg puncta with LysoTracker endolysosomes by Pearson’s correlation coefficient. (C and C′) Methylarginine and BSA-DQ lysosomal marker (arrowheads) colocalize with Wnt3a. Panel C′ was treated with Wnt3a for 15 min. (D) Wnt increased colocalization of meArg puncta with BSA-DQ endolysosomes by Pearson’s correlation coefficient. (E) Wnt signaling (15 min) increased endocytosis of BSA-FITC and BSA-DQ into the cell liquid phase by quantification of fluorescence per cell; n > 20 cells per sample. (F and G) Wnt signaling increased BSA-FITC and BSA-DQ uptake by flow cytometry (4-h treatment) (36). (H) Blocking protein synthesis with cycloheximide did not prevent Wnt-induced BSA-DQ endocytosis. (I–K) Wnt-induced meArg sequestration into MVBs marked by Vps4-GFP (arrowheads) was blocked by DN-Vps4. (L–N) Lys48-polyUb proteins colocalized with Vps4-GFP MVBs (arrowheads) 15 min after Wnt3a treatment and this required active Vps4. (Scale bars, 10 μm.) *P < 0.05.