Figure 2: A lysosome-targeted genetically encoded Ca2+ indicator. | Nature Communications

Figure 2: A lysosome-targeted genetically encoded Ca2+ indicator.

From: Lipid storage disorders block lysosomal trafficking by inhibiting a TRP channel and lysosomal calcium release

Figure 2

(a) GCaMP3–TRPML1 (GCaMP3–ML1) fusion strategy. GCaMP3 is fused to the N-terminus of TRPML1. (b) Colocalization of GCaMP3-ML1 fluorescence with Lamp-1 in HEK293T cells expressing both GCaMP3-ML1 and Lamp-1-mCherry. Scale bar=5 μm. (c) Colocalization of GCaMP3-ML1 fluorescence with LysoTracker in GCaMP3-ML1-expressing HEK293T cells. Scale bar=2 μm. (d) Preferential detection of Ca2+ release from lysosome stores by GCaMP3-ML1 in GCaMP3-ML1-transfected Cos-1 cells. ER and lysosome Ca2+ releases were triggered by thapsigargin (TG, 2 μM) and GPN (200 μM), respectively, in low (nominally free Ca2+ + 1 mM EGTA; free [Ca2+] estimated to be <10 nM) external [Ca2+]. All cells responded to GPN; only one out of four cells responded to TG.

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