Journal: bioRxiv

Article Title: A small molecule cocktail for robust induction of blood-brain barrier properties

doi: 10.1101/2023.02.09.527899

Figure Lengend Snippet: a) Schematic drawing of the experimental setup. MACE-seq was performed on samples from brain-like ECs (in co-culture with PCs), with or without 48 h cARLA treatment. b) Principal component analysis (PCA) plot of control and cARLA samples. c) Venn-diagram showing the number of genes upregulated (log 2 FC ≥ 0.2 and FDR < 0.01), downregulated (log 2 FC ≥ -0.2 and FDR < 0.01) or not changed (|log 2 FC| < 0.2 and/or FDR ≥ 0.01) upon cARLA treatment. Log 2 FC: log 2 (fold change), FDR: false discovery rate. d) Mean-difference (MD) plot presenting the range and distribution of all transcripts (circles) regarding their mean expression and log 2 FC upon cARLA treatment. Key transcripts related to different aspects of BBB function are highlighted. e) Functional enrichment analysis of up- and downregulated gene sets upon cARLA treatment using g:Profiler. Gene Ontology Biological Process (GO:BP) terms are ranked based on their gene ratio (the fraction of differentially expressed genes in a given term) and are colored based on statistical significance (FDR). f) Representative staining of F-actin (grey), with or without cARLA treatment. Nuclei were stained with Hoechst (blue). Bar: 40 µm. g) Validation of the definitive endothelial nature of human stem cell-derived brain-like ECs. Normalized counts of key epithelial and endothelial transcripts from our dataset are plotted on a 3-segment y-axis covering multiple orders of magnitude. h) Scaled heatmap of 92 transcripts that are enriched in mouse brain ECs and are well expressed at the human BBB. Genes upregulated, downregulated or not changed upon cARLA treatment as well as genes not expressed (normalized counts ≤ 1) in this model are shown separately, in alphabetical order. Color coding represents row z-scores after Min-Max normalization, the darkest color corresponding to the highest expression in a given row.

Article Snippet: A total of 8 libraries (4 control and 4 cARLA) were constructed using the Rapid MACE-Seq kit (GenXPro GmbH, Frankfurt, Germany) according to the manufacturer’s protocol.

Techniques: Co-Culture Assay, Expressing, Functional Assay, Staining, Derivative Assay

Journal: bioRxiv

Article Title: A small molecule cocktail for robust induction of blood-brain barrier properties

doi: 10.1101/2023.02.09.527899

Figure Lengend Snippet: Gene expressional changes after 48 h cARLA treatment were determined using MACE-seq profiling and are presented in panels a-i) by category. Mean ± SD, n =4. Red and blue color indicates up- and downregulation, respectively, upon cARLA treatment.

Article Snippet: A total of 8 libraries (4 control and 4 cARLA) were constructed using the Rapid MACE-Seq kit (GenXPro GmbH, Frankfurt, Germany) according to the manufacturer’s protocol.

Techniques: