Fig. 1: A novel Shank3 variant is expressed in BECs.
a Collection of cDNAs from RNA extracts of mouse primary BECs, bEnd.3 BECs, and primary neurons. Created in BioRender. Kim, S. (2025) https://BioRender.com/q10o614. b PCR analysis revealed that mouse primary BECs and bEnd.3 BECs express only Shank3 mRNA, while neurons express all Shank family. β-Actin: input control. Created in BioRender. Kim, S. (2025) https://BioRender.com/c22u541. c Sequencing chromatograms showing the lack of exon 18 in Shank3 mRNA in mouse primary BECs and bEnd.3 BECs, whereas neuronal Shank3 has exon 18. d PCR analysis confirmed exon 18-deleted Shank3 in the mouse primary BECs and bEnd.3 BECs. Shank3Δe18: exon 18 deletion form of Shank3. PCR (e) and qRT-PCR (f) analyses demonstrated that neonatal Shank3 mRNA expression at P5 (n = 4) was significantly higher than that in adult mice (n = 3), in both males and females. The graph represents the relative expression of Shank3 mRNA normalized to its levels in P5 male mice. Gapdh (Glyceraldehyde-3-phosphate dehydrogenase): input control. g ICC analysis revealed that eSHANK3 is mainly expressed in ZO1-positive cell-cell junctional area of mouse primary BECs and bEnd.3 BECs. Insets showing magnified views of the junctional areas. DAPI: nuclear staining. Scale bar, 20 μm. Each experiment b, d, g was independently repeated three times, yielding similar results. Data are mean ± SEM. Statistical analysis was performed using two-way ANOVA followed by Šídák’s multiple comparisons test (f). Detailed statistical methods and results are described in Supplementary Table 1. Source data are provided as Source Data File.
