Research Paper Volume 13, Issue 12 pp 15917—15941

Figure 1. G4 landscapes vary among primary neurons, astrocytes, and microglia. (A) Primary cortical neurons (14 DIV) were fixed and stained with 25 μM N-TASQ, antibodies against MAP2c, and the nuclear dye DAPI, and imaged with a confocal microscope. Note the N-TASQ-positive structure in the nucleus of the cell on the left (depicted with arrow). Scale bar, 5 μm. (B) Primary cortical cultures were stained and imaged as in (A). Note the MAP2c-positive neurons on the right that contain small N-TASQ-positive puncta in the cytoplasm (depicted with arrow). Note the MAP2c-negative cell on the right that contains many N-TASQ-positive puncta in the cytoplasm (depicted with arrow). Scale bar, 10 μm. (C) Cultured primary astrocytes were fixed, stained with 25 μM N-TASQ, antibodies against GFAP, and DAPI, and imaged with a confocal microscope. Note numerous N-TASQ-positive structures in the nucleus and cytoplasm. Scale bar, 10 μm. (D) Cultured primary microglial cells were fixed, stained with 25 μM N-TASQ, antibodies against Iba-1, and DAPI, and imaged with a confocal microscope. Note N-TASQ-positive structures in the nuclei and the cytoplasm (depicted with arrows). Scale bar, 5 μm. (E) The nuclear puncta index of the N-TASQ staining was analyzed in the nuclei of cells from (A), (C, D). The Kruskal-Wallis test was used. ****p<0.0001. 120 cells per cell type were analyzed, and results were pooled from three independent experiments. (F) The cytoplasmic puncta index of the N-TASQ staining was analyzed in the cytoplasm of cells from (A), (C, D). The Kruskal-Wallis test was used. ****p<0.0001, ***p=0.0001. 120 cells per cell type were analyzed, and results were pooled from three independent experiments.