Extended Data Fig. 4: GABAergic neuron-to-glioma synaptic puncta imaging and 3D reconstruction.
From: GABAergic neuron-to-glioma synapses in diffuse midline gliomas
a. 3-dimensional (3D) reconstruction shown in Fig. 2c and associated zoomed-out confocal images of GABAergic neuron-to-glioma synaptic puncta in neuron-glioma co-cultures. H3K27M+ DMG cells (SU-DIPGXIII-FL, nestin, blue) expressing GARBRG2-GFP (green, post-synaptic) co-localizes with synapsin (white, pre-synaptic) on rat hippocampal neurons (neurofilament, red). Scale bars, 2 µm. b. Zoomed-out 3-dimensional (3D) reconstructions and zoomed-in 3D reconstructions (as shown in Fig. 2d,e) with associated confocal images of GABAergic neuron-to-glioma synaptic puncta in patient-derived DMG xenografts in the mouse hippocampus. Gephyrin (green) on GFP-expressing DMG cells (SU-DIPGXIII-FL, GFP,blue) co-localizes with VGAT (white) on neurons (DLX-mCherry, red). Scale bars, 2 µm. Top left, quantification of co-localized synaptic puncta as a percent of total gephyrin puncta on glioma cells (n = 4 mice). Select colocalized puncta are indicated with yellow arrows, and some examples are highlighted in yellow boxes. Please note that several 3D-reconstructed images from Fig. 2 are shown again here to provide additional spatial and contextual information. All data are mean ± s.e.m.
