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. 2022 Sep 5;5:908. doi: 10.1038/s42003-022-03878-9

Fig. 2. Knockdown of elevated CPEB2 Expression in GECs Increases BTB Permeability.

Fig. 2

a Venn diagram. (Group1, MeRIP-seq of scramble and METTL3 knockdown HEK-293T cell, GSE182607; Group2, Transcriptome analysis of gene expression in scramble and METTL3 knockdown HUVEC (human umbilical vein endothelial cells) cell lines, GSE157544; Group 3, RNA deep sequencing analysis of human brain microvascular endothelial cells (ECs) treated with glioma-conditioned medium (glioma-CM), GSE115850; Group 4, Genes positively associated with poor prognosis in glioma.) b Correlation heatmap of METTL3 and IGF2BP3 with differentially expressed factors in glioma, analyzed using TCGA database. c Venn diagram. d Relative mRNA levels of KLF6, CPEB2, and TMED9 in AECs and GECs were determined by qRT-PCR. Data are represented as mean ± SD (n = 3). **P < 0.01 vs. AECs group. e Relative mRNA levels of KLF6, CPEB2, and TMED9 in sh-METTL3 GECs were determined by qRT-PCR. Data are represented as mean ± SD (n = 3). **P < 0.01 vs. sh-NC group. f RIP was performed with normal mouse IgG or anti-IGF2BP3 antibody in GECs. Data are represented as mean ± SD (n = 3). **P < 0.01 vs. anti-IgG group. Relative enrichment of KLF6, CPEB2, and TMED9 was determined by qRT-PCR. g CPEB2 prognostic analysis in glioma using TCGA database. h Relative protein levels of CPEB2 in AECs and GECs were determined by western blot analysis. i, j The permeability and integrity of the CPEB2 knockdown in vitro BTB model were detected by TEER values, FITC-dextran, and HRP flux. Data are represented as mean ± SD (n = 5). **P < 0.01 vs. sh-NC group. k The expressions of ZO-1, occludin, and claudin-5 in the CPEB2 knockdown GECs detected by western blot analysis. Data are represented as mean ± SD (n = 3). **P < 0.01 vs. sh-NC group. l The distributions of ZO-1, occludin, and claudin-5 in the CPEB2 knockdown GECs were observed by IF staining. Scale bar represents 50 µm.