Fig. 4
From: Sub-nanomolar sensitive GZnP3 reveals TRPML1-mediated neuronal Zn2+ signals
Target Zn2+ sensor to subcellular compartments to detect local TRPML1-mediated signals. a Schematic of protein constructs for (1) GZnP3-TRPML1, (2) LAMP1-GZnP3, (3) GZnP3-Rab7a, and (4) synaptophysin-GZnP3 (syn-GZnP3) with N- and C-termini marked (left). Diagram of subcellular localizations of each targeted GZnP3 construct (right). b Representative traces of HeLa cells expressing GZnP3 (gray), GZnP3-TRPML1 (green), LAMP1-GZnP3 (black) GZnP3-Rab7a (magenta), or synaptophysin-GZnP3 (blue). Cells were treated with 100 µM TPEN at 0 s (white arrow), washed (black bar), then treated with 100 µM ZnCl2 and 2.5 µM pyrithione at 300 s (black arrow). c Mean sensor dynamic range (±s.e.m.) of cytosolic control GZnP3 (gray, n = 13 cells) and vesicular-targeted variants: GZnP3-TRPML1 (green, n = 23), LAMP1-GZnP3 (black, n = 10), GZnP3-Rab7a (magenta, n = 12), and synaptophysin-GZnP3 (blue, n = 13). HeLa cells were treated as described in b. Bars indicate mean fold-change of sensor fluorescence (Fmax/Fmin). One-way ANOVA with Dunnett’s multiple comparison to GZnP3. ****p < 0.0001, n.s. not significant. p-values listed in italics above corresponding comparisons where applicable. Source data are provided as a Source Data file
