Journal: EMBO Reports

Article Title: miRNA-target complementarity in cnidarians resembles its counterpart in plants

doi: 10.1038/s44319-024-00350-z

Figure Lengend Snippet: Reagents and tools table

Article Snippet: In vitro transcription was conducted with HighYield T7 Cap 1 AG (3‘-OMe) mRNA Synthesis Kit (m5CTP) (Jena Bioscience, Germany) using 800 ng of amplified template followed by Turbo DNase treatment (Thermo Fisher Scientific) by incubation with 1 µl of DNase for 30 min at 37 °C twice sequentially.

Techniques: Sequencing, shRNA, SYBR Green Assay, Protease Inhibitor, Bicinchoninic Acid Protein Assay, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Magnetic Beads, Multiplex Assay, Sensitive Assay, Software, Imaging, Spectrophotometry, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: Liver-Specific Overexpression of Prostasin Attenuates High-Fat Diet-Induced Metabolic Dysregulation in Mice

doi: 10.3390/ijms22158314

Figure Lengend Snippet: Serum PRSS8 levels was negatively associated with the presence of type 2 diabetes and increased intima media thickness. ( a ) The ELISA of human PRSS8 levels in the serum of the nondiabetes mellitus (non-DM) and type 2 diabetes mellitus (T2DM) participants (n = 39–40 per group). Values are shown as the mean ± SEM; * p -value < 0.05 ( t test). ( b ) The maximum intima media thickness (max IMT) measured by carotid ultrasonography in T2DM. Values are shown as the mean ± SEM; ** p -value < 0.01 ( t test). ( c ) Homeostatic model assessment beta cell function (HOMA-β) in non-DM was shown (Pearson product-moment correlation coefficient).

Article Snippet: Measurement of serum fasting PRSS8 levels in WT/LTg mice was used by Human Prostasin ELISA Kit (BOSTER Biological Technology Co., Ltd. Pleasanton, CA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Cell Function Assay

Journal: Biology Direct

Article Title: CAP1: a novel extracellular vesicle marker linked to endothelial senescence in atherosclerosis

doi: 10.1186/s13062-025-00646-7

Figure Lengend Snippet: CAP1 induces ES in HAEC and CAEC. (A) Representative immunoblot detection of CAP1 in HAEC and CAEC from P1 to P12. ( N = 3 Mean ± SD. * p < 0.05. # p < 0.01. $ p < 0.001 P1 vs. selected passages). (B) Representative immunoblot detection of CAP1 in conditioned medium of HAEC and CAEC from P1 to P12. ( N = 3 Mean ± SD. * p < 0.05. P1 vs. selected passages). (C) Senescence-associated Β-gal (SABG) staining of P1 HAEC and CAEC incubated 24 h with 2 ml culture medium from P1-12 HAEC and CAEC as indicated ( N = 3 Mean ± SD. * p < 0.05. ** p < 0.01. P1 vs. selected passages). (D) CAP1 silenced HAEC and CAEC representative immunoblot shows. N = 3 Mean ± SD. * p < 0.05 CAP1 siRNA vs. CONTROL. (E) CAP1 silenced HAEC and CAEC overexpressing CAP1 (after transfection of pCAP1 ), as shown by representative immunoblot. N = 3 Mean ± SD. * p < 0.05 CAP1 siRNA vs. CAP1 siRNA/pCAP1. (F) Representative immunoblot detection of PCSK9 in HAEC and CAEC from P1 to P12. ( N = 3 Mean ± SD). (G) ES assay in HAEC and CAEC incubated with 100 mM PCSK9 inhibitor Evolocumab. N = 3 Mean ± SD. (H) ES assay in Evolocumab incubated cells as in B. N = 3 Mean ± SD, * p < 0.05, and # p < 0.05 CONTROL vs. CAP1 siRNA in HAEC and CAEC, respectively. @ p < 0.05 HAEC CAP1 siRNA/pCAP1 and CAP1 siRNA/pCAP1/Evo vs. CONTROL. $ p < 0.05 CAEC CAP1 siRNA/pCAP1 and CAP1 siRNA/pCAP1/Evo vs. CONTROL

Article Snippet: HAEC and CAEC were seeded in 35 mm glass-bottom culture dishes and incubated for 24 h with Lipofectamine RNAiMAX transfection reagent (Thermo Fisher) containing 1 µg/plasmid DNA (CAP1 (NM_001105530)) (Origene) in medium supplemented with neomycin (500 µg/ml).

Techniques: Western Blot, Staining, Incubation, Control, Transfection

Journal: Biology Direct

Article Title: CAP1: a novel extracellular vesicle marker linked to endothelial senescence in atherosclerosis

doi: 10.1186/s13062-025-00646-7

Figure Lengend Snippet: Expression of CAP1 in mouse and human atherosclerosis. (A) Representative immunohistochemistry detection of CAP1 in aortic plaques from young and aged ApoE atherosclerotic mice. Scale bars 50 μm. N = 3 mice/group. * p < 0.05 Young vs. Aged ApoE KO mice. (B) Representative immunohistochemistry detection of CAP1 in human carotid endarterectomy sections from group 1 (patients younger than 60 years, upper panels) and group 2 (patients older than 80 years, lower panels). Scale bars 100 μm. N = 3 patients/group. P < 0.005 group1 vs. group2

Article Snippet: HAEC and CAEC were seeded in 35 mm glass-bottom culture dishes and incubated for 24 h with Lipofectamine RNAiMAX transfection reagent (Thermo Fisher) containing 1 µg/plasmid DNA (CAP1 (NM_001105530)) (Origene) in medium supplemented with neomycin (500 µg/ml).

Techniques: Expressing, Immunohistochemistry

Journal: Biology Direct

Article Title: CAP1: a novel extracellular vesicle marker linked to endothelial senescence in atherosclerosis

doi: 10.1186/s13062-025-00646-7

Figure Lengend Snippet: EVs from aged ApoE knockout mice induce ES in WT animals. A. Representative immunoblot detection of CAP1 from blood circulating EVs isolated from young and aged ApoE knockout mice ( N = 3 Mean ± SD. * p < 0.05 Young vs. Aged ApoE EVs). B-E. Hematoxylin/Eosin (H&E) staining (upper panels) and b-Galactosidase (b-Gal) assay (lower panels) in aortic sections of WT mice fed with regular diet ( B , C ) or high-fat diet ( D , E ), and injected with EVs from young ( B , D ), or aged ( C , E ) ApoE KO mice as indicated. Scale bars 100 μm ( N = 3/group)

Article Snippet: HAEC and CAEC were seeded in 35 mm glass-bottom culture dishes and incubated for 24 h with Lipofectamine RNAiMAX transfection reagent (Thermo Fisher) containing 1 µg/plasmid DNA (CAP1 (NM_001105530)) (Origene) in medium supplemented with neomycin (500 µg/ml).

Techniques: Knock-Out, Western Blot, Isolation, Staining, Injection