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Histological localization of PUM1 in brain tissue from patients with epilepsy. ( a to f ) The expression level of CLDN-5 mRNA in resected human brain samples. The RNAs were isolated from the resected brain tissues obtained from control subjects ( n = 11) and patients with epilepsy ( n = 12). ( a ) CLDN-5, PUM1 and PUM2 mRNA expression levels between samples collected from control and epilepsy patients. CLDN-5 mRNA levels in the samples were broken down by ( b ) the difference of rs10314 genotypes (G is wild-type allele and C is rs10314 allele) and ( c ) seizure frequency. The data represents mean ± SD. ns, not significant ( p > 0.05); *p value ≤ 0.05; **p value ≤ 0.01. The correlation between CLDN-5 mRNA and ( d ) years of epilepsy, ( e ) PUM1 and ( f ) PUM2 mRNA levels. Spearman r values and P values are indicated. ( g ) Representative confocal images of human brain sections stained with anti-PUM1 antibody. A part of the hippocampal region showed a granular PUM1 staining pattern. Red; PUM1, Green; CLDN-5. Blue; nuclei. ( h ) The evaluation of the induction of stress granules in vascular endothelial cells in human brain sections (high CLDN-5). Sections were stained with anti-CLDN-5 (green) and stress granule marker, <t>anti-p-eIF-2α</t> or anti-PUM1 (red). Nuclei (blue) were counterstained with Hoechst
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Histological localization of PUM1 in brain tissue from patients with epilepsy. ( a to f ) The expression level of CLDN-5 mRNA in resected human brain samples. The RNAs were isolated from the resected brain tissues obtained from control subjects ( n = 11) and patients with epilepsy ( n = 12). ( a ) CLDN-5, PUM1 and PUM2 mRNA expression levels between samples collected from control and epilepsy patients. CLDN-5 mRNA levels in the samples were broken down by ( b ) the difference of rs10314 genotypes (G is wild-type allele and C is rs10314 allele) and ( c ) seizure frequency. The data represents mean ± SD. ns, not significant ( p > 0.05); *p value ≤ 0.05; **p value ≤ 0.01. The correlation between CLDN-5 mRNA and ( d ) years of epilepsy, ( e ) PUM1 and ( f ) PUM2 mRNA levels. Spearman r values and P values are indicated. ( g ) Representative confocal images of human brain sections stained with anti-PUM1 antibody. A part of the hippocampal region showed a granular PUM1 staining pattern. Red; PUM1, Green; CLDN-5. Blue; nuclei. ( h ) The evaluation of the induction of stress granules in vascular endothelial cells in human brain sections (high CLDN-5). Sections were stained with anti-CLDN-5 (green) and stress granule marker, <t>anti-p-eIF-2α</t> or anti-PUM1 (red). Nuclei (blue) were counterstained with Hoechst
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Histological localization of PUM1 in brain tissue from patients with epilepsy. ( a to f ) The expression level of CLDN-5 mRNA in resected human brain samples. The RNAs were isolated from the resected brain tissues obtained from control subjects ( n = 11) and patients with epilepsy ( n = 12). ( a ) CLDN-5, PUM1 and PUM2 mRNA expression levels between samples collected from control and epilepsy patients. CLDN-5 mRNA levels in the samples were broken down by ( b ) the difference of rs10314 genotypes (G is wild-type allele and C is rs10314 allele) and ( c ) seizure frequency. The data represents mean ± SD. ns, not significant ( p > 0.05); *p value ≤ 0.05; **p value ≤ 0.01. The correlation between CLDN-5 mRNA and ( d ) years of epilepsy, ( e ) PUM1 and ( f ) PUM2 mRNA levels. Spearman r values and P values are indicated. ( g ) Representative confocal images of human brain sections stained with anti-PUM1 antibody. A part of the hippocampal region showed a granular PUM1 staining pattern. Red; PUM1, Green; CLDN-5. Blue; nuclei. ( h ) The evaluation of the induction of stress granules in vascular endothelial cells in human brain sections (high CLDN-5). Sections were stained with anti-CLDN-5 (green) and stress granule marker, <t>anti-p-eIF-2α</t> or anti-PUM1 (red). Nuclei (blue) were counterstained with Hoechst
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Histological localization of PUM1 in brain tissue from patients with epilepsy. ( a to f ) The expression level of CLDN-5 mRNA in resected human brain samples. The RNAs were isolated from the resected brain tissues obtained from control subjects ( n = 11) and patients with epilepsy ( n = 12). ( a ) CLDN-5, PUM1 and PUM2 mRNA expression levels between samples collected from control and epilepsy patients. CLDN-5 mRNA levels in the samples were broken down by ( b ) the difference of rs10314 genotypes (G is wild-type allele and C is rs10314 allele) and ( c ) seizure frequency. The data represents mean ± SD. ns, not significant ( p > 0.05); *p value ≤ 0.05; **p value ≤ 0.01. The correlation between CLDN-5 mRNA and ( d ) years of epilepsy, ( e ) PUM1 and ( f ) PUM2 mRNA levels. Spearman r values and P values are indicated. ( g ) Representative confocal images of human brain sections stained with anti-PUM1 antibody. A part of the hippocampal region showed a granular PUM1 staining pattern. Red; PUM1, Green; CLDN-5. Blue; nuclei. ( h ) The evaluation of the induction of stress granules in vascular endothelial cells in human brain sections (high CLDN-5). Sections were stained with anti-CLDN-5 (green) and stress granule marker, <t>anti-p-eIF-2α</t> or anti-PUM1 (red). Nuclei (blue) were counterstained with Hoechst
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Histological localization of PUM1 in brain tissue from patients with epilepsy. ( a to f ) The expression level of CLDN-5 mRNA in resected human brain samples. The RNAs were isolated from the resected brain tissues obtained from control subjects ( n = 11) and patients with epilepsy ( n = 12). ( a ) CLDN-5, PUM1 and PUM2 mRNA expression levels between samples collected from control and epilepsy patients. CLDN-5 mRNA levels in the samples were broken down by ( b ) the difference of rs10314 genotypes (G is wild-type allele and C is rs10314 allele) and ( c ) seizure frequency. The data represents mean ± SD. ns, not significant ( p > 0.05); *p value ≤ 0.05; **p value ≤ 0.01. The correlation between CLDN-5 mRNA and ( d ) years of epilepsy, ( e ) PUM1 and ( f ) PUM2 mRNA levels. Spearman r values and P values are indicated. ( g ) Representative confocal images of human brain sections stained with anti-PUM1 antibody. A part of the hippocampal region showed a granular PUM1 staining pattern. Red; PUM1, Green; CLDN-5. Blue; nuclei. ( h ) The evaluation of the induction of stress granules in vascular endothelial cells in human brain sections (high CLDN-5). Sections were stained with anti-CLDN-5 (green) and stress granule marker, anti-p-eIF-2α or anti-PUM1 (red). Nuclei (blue) were counterstained with Hoechst

Journal: Fluids and Barriers of the CNS

Article Title: Pumilio-1 mediated translational control of claudin-5 at the blood-brain barrier

doi: 10.1186/s12987-024-00553-5

Figure Lengend Snippet: Histological localization of PUM1 in brain tissue from patients with epilepsy. ( a to f ) The expression level of CLDN-5 mRNA in resected human brain samples. The RNAs were isolated from the resected brain tissues obtained from control subjects ( n = 11) and patients with epilepsy ( n = 12). ( a ) CLDN-5, PUM1 and PUM2 mRNA expression levels between samples collected from control and epilepsy patients. CLDN-5 mRNA levels in the samples were broken down by ( b ) the difference of rs10314 genotypes (G is wild-type allele and C is rs10314 allele) and ( c ) seizure frequency. The data represents mean ± SD. ns, not significant ( p > 0.05); *p value ≤ 0.05; **p value ≤ 0.01. The correlation between CLDN-5 mRNA and ( d ) years of epilepsy, ( e ) PUM1 and ( f ) PUM2 mRNA levels. Spearman r values and P values are indicated. ( g ) Representative confocal images of human brain sections stained with anti-PUM1 antibody. A part of the hippocampal region showed a granular PUM1 staining pattern. Red; PUM1, Green; CLDN-5. Blue; nuclei. ( h ) The evaluation of the induction of stress granules in vascular endothelial cells in human brain sections (high CLDN-5). Sections were stained with anti-CLDN-5 (green) and stress granule marker, anti-p-eIF-2α or anti-PUM1 (red). Nuclei (blue) were counterstained with Hoechst

Article Snippet: The sections were incubated with mouse anti-CLDN-5 antibodies (clone 4C3C2, Thermo Fisher Scientific) and rabbit anti-PUM1 antibodies (clone EPR3795, Abcam) or rabbit anti- p-eIF-2α antibodies (44-728G, Thermo Fisher Scientific) in 1% NGS overnight at 4 °C.

Techniques: Expressing, Isolation, Staining, Marker