Fig. 2

22L-derived reactive astrocytes show impairment in supporting neuronal growth. Primary cortical neurons were plated on CT-PACs or 22L-PACs and co-cultured for 10–12 days. a Representative fluorescent images of neuron-astrocyte co-cultures co-immunostained for MAP2 (green) and GFAP (red), and quantification of neurite length. b Representative fluorescent images of cortical neuronal cells co-cultured with CT- PACs or 22L-PACs, and co-immunostained for pre- and post-synaptic markers synaptophysin (SYP, green) and PSD95 (red), respectively, and MAP2 (blue). Co-localization between SYP and PSD95 was analyzed for quantification of synapses in neuronal cells co-cultured with CT-PACs or 22L-PACs (yellow puncta). c Representative images of cortical neuronal cells co-cultured with CT-PACs or 22L-PACs, and co-immunostained with a spine marker Drebrin (red) and MAP2 (green). Quantification of spine size and density in neurons co-cultured with CT-PACs and 22L-PACs. In a–c, data represent mean ± SE, n = 3 independent experiments in which CT-PACs or 22L-PACs were isolated individual animals, 50 neurons (in a and b) and 40 neurons (in c) per conditions were analyzed, **p < 0.01 and *p < 0.05 (two tailed, unpaired t test with Welch's correction). Scale bars = 50 µm in a, 25 µm in b, and 10 µm in c