Fig 1: Agrin and laminin improve NMJ formation in microfluidic devices(A and B) Confocal micrographs of NMJs in agrin (0.01 μg/mL)- and laminin (20 μg/mL)-supplemented (A/L) and untreated (Con) conditions on day 28 in XC150 devices. Single contact point NMJs (A). Multiple contact point NMJs (B). Scale bars, 10 μm. Arrowheads: Btx-SYP/NEFH co-localizations.(C) Number of Btx and SYP/NEFH co-localizations per myotube.(D) Quantifications of NMJ single and multiple contact point morphology.(E and F) Representative scanning electron microscopy (SEM) images of NMJ formation in SND75 devices on day 28 in A/L-supplemented and control conditions. Single contact point NMJs (E). Multiple contact point NMJs (F). Insets: magnifications of NMJ (arrowheads). Scale bars, 2 μm and 1 μm (insets). (C) Unpaired t test. (D) One-way ANOVA with Tukey's multiple comparisons test. Mean ± SEM of four biological replicates. ∗p < 0.05, ∗∗p < 0.01.See also Figures S2 and S3.
Fig 2: Agrin promotes the development of SE-induced TLE in an Lrp4-dependent manner. A Experimental design for SRS recording. KA was infused into the amygdala of C57 wildtype mice to induce SE, which was terminated one hour later by injection of diazepam (i.p). During the latent period of 2 weeks, agrin was infused into the lateral ventricle each day. SRS were then recorded for 1 week. B Increased SRS number in Agrin-treatment group. n = 10 mice for each group. Student’s t test, t(18) = 3.428, p = 0.003. C Simlar duration of SRS between the two groups. n = 10 mice for each group. Student’s t test, t(18) = 0.429, p = 0.1701. D Simlar seizure stage between the two groups. n = 10 mice for each group. Student’s t test, t(18) = 0.021, p = 0.9835. E Experimental design. Virus was bilaterally injected into the hippocampus of Lrp4f/f mice and a guide canula was implanted in the right amygdala. 3 days later, KA was infused into the amygdala to induce SE, which was terminated one hour later by injection of diazepam (i.p). During the latent period of 2 weeks, agrin was infused into the lateral ventricle each day. SRS were then recorded for 1 week. F Diminished change in SRS number in hGFAP-Cre group by Agrin-treatment. n = 6 mice for ACSF group; n = 5 mice for agirn group. Student’s t test, t(9) = 1.933, p = 0.0853. G Simlar duration of SRS between the two groups. n = 6 mice for ACSF group; n = 5 mice for agirn group. Student’s t test, t(9) = 0.6621, p = 0.5245. H Simlar SRS stage between the two groups. n = 6 mice for ACSF group; n = 5 mice for agirn group. Student’s t test, t(9) = 0.1574, p = 0.8784
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