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. 2021 Jun 15;131(12):e140723. doi: 10.1172/JCI140723

Figure 4. Quantitation of G3BP1 SGs using the Imaris 3D masking method.

Figure 4

(A) SGs were induced in vitro with SA treatment, and detected by immunofluorescence with an antibody against G3BP1 (green). The Imaris surface rendering tool was used to detect G3BP1 SG puncta (gray). Scale bar: 10 μm. (B) Using in vitro–induced SGs, parameters were set for the detection of G3BP1 granules in brain tissue. Z stacks were obtained at a thickness of 0.5 μm per slice, and analyzed using the surface rendering tool for quantitation of G3BP1 puncta. The number of puncta was normalized to the number of nuclei (DAPI) per frame. (C) The same pipeline was applied to detect G3BP1 granules in brain tissue, allowing for the quantitation of G3BP1 puncta and not diffuse G3BP1 background signal. Original magnification ×100.