Fig. 2: Recapitulated NsMJs in vitro.
a, Bright-field and fluorescence microscopy images of the primary neurons and the tdTomato+ aSMCs, showing a putative NsMJ. b, The orange lollipop plot represents a single quantification for the example in a. The insert shows the percentages of cells that formed stable contacts with neurite terminals in the total tdTomato– cells (N = 145 cells) or the total tdTomato+ cells (N = 150 cells; n = 14 field of views from 7 dishes of co-cultures). Data are the mean ± s.e.m.; nested, unpaired, two-tailed t-test. c,d, SEM of axons and aSMCs that protrude microvilli. e,f, CLEM of co-culture. SEM image (e) correlates with the confocal image (f), labeled with anti-Tau and anti-α-SMA. g, TEM image of the ultrastructure of an NsMJ on a tdTomato+ aSMC in th e insert labeled by the magenta arrow (Supplementary Fig. 6). The axonal bouton, ECM, aSMC cell membrane and aSMC are highlighted in light green, yellow, blue dashed lines and red. h, The thickness of aSMC ECM flanking NsMJs (n = 5 NsMJs, depicted by each shape of symbols). i, The correlation of ECM thickness of aSMC with the distance flanking NsMJs (n = 5 NsMJs). j, Fluorescence microscopy image of CTBAlex-488 of the co-culture at day 7 in vitro. Neuronal soma (red arrow); CTBAlex488-pretreated HB-vSMCs (white dashed line); CTBAlex488-positive axon (green arrows). The white frame with magenta arrow in the insert shows higher magnification of the NsMJ location. k, Pie graph of percentages of CTBAlex488-positive or negative neurons in co-cultured cortical neurons from three independent co-culture replicates.
