HEPACAM Antibody - #DF12075

Fig. 2 Involvement of A-Exo. surface signals in promoting axon growth and identification of the surface expression of HepaCAM (GlialCAM) on A-Exo. Representative images (a) and quantification (b) of axon length of cortical neurons in control (i) or treated with proteinase K (10 μg/ml, 5 min) digested A-Exo. (ii) sonicated (30 s) and proteinase K digested A-Exo. (iii) A-Exo. (iv) sonicated A-Exo. (v) or sonicated (30 s) and RNase (10 μg/ml, 5 min) digested A-Exo. (vi) 1 μg A-Exo./sample was used in each treatment in (a, b). White arrows: elongated axons; Number of neurons quantified in each group shown in the graph (6–11 neurons/replicate, 3 biological replicates)/group; Scale bar: 100 μm; Representative images (c) and quantification (d) of axon length of cortical neurons plated on either poly-D-lysine (PDL) coated or PDL/A-Exo. coated coverslips. Number of neurons quantified in each group shown in the graph (10–11 neurons/replicate, 3 biological replicates)/group; Scale bar: 100 μm; e Quantification of axon length of cortical neurons following A-Exo. treatment or co-treatment with A-Exo. and dynasore (dynamin inhibitor, 50 μM). n = 22 (control) or 21 (A-Exo. and A-Exo. + dynasore) neurons (6–7 neurons/replicate, 3 biological replicates)/group; f Proteomic identification of different categories of transmembrane proteins on A-Exo. surface. Specific transmembrane proteins are included in Supplementary Table 1. n = 3 biological replicates; g Detection of specific HepaCAM immunoreactivity (>5 replicates) from spinal cord lysate (10 μg/lane), astrocyte lysate (10 μg/lane), and A-Exo. (1 μg/lane) prepared from WT (+/+), HepaCAM heterozygous (+/−), and HepaCAM KO (−/−) mice; Red arrow: specific HepaCAM immunoreactivity; Black arrow: non-specific immunoreactivity; h Detection of specific HepaCAM immunoreactivity (>5 replicates) in A-Exo. but not in exosome-free ACM fractions; 1 μg A-Exo. was used in each experiment. p value in (d) determined from two-tailed t-test; p values in (b, e) determined by one-way ANOVA followed by post hoc Tukey’s test. Data are presented as mean values ± SEM.