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. 2016 Oct 18;11(3):196–208. doi: 10.1080/19336950.2016.1249076

Figure 6.

Figure 6.

TRPV3 is present in lysosomes and OS-mutants alter lysosomal numbers and movement. (A) Time-series confocal images were acquired for analyzing lysosomal movements in stable cells expressing TRPV3-Wt –GFP or OS-mutants. DIC images (white dotted lines show the periphery of the cells) superimposed with the florescence images (for Lysotraker-red labeled Lysosomes, red circles). The track-index of Lysosomal movements are indicated by lines (Right panel). For more details see movies (S1-S5). (B) HaCaT cells stably expressing OS-mutants have lower number of lysosomes (labeled with Lysotracker red) than the cells expressing TRPV3-Wt-GFP (P-values: *** < 0.001, ** < 0.01 TRPV3-G573C or TRPV3-G573S; n = 20 cells in each case). (C) Confocal images of stable HaCaT cells show that both TRPV3-Wt-GFP and OS-mutants:localize (white arrows) in lysosomes in live cells. Colocalization experiments with TRPV3-G573A and TRPV3-W692G failed mainly due to the undetectable level of tagged proteins and absence of lysosomes properly labeled with Lysotracker red. (D) Western blot analysis of lysosomal fraction isolated from goat brain shows the presence of endogenous TRPV3 there (indicated by arrow) and enrichment of lysosomal fraction shown with lysosomal marker.