Journal: Diabetes

Article Title: Hsp20-Mediated Activation of Exosome Biogenesis in Cardiomyocytes Improves Cardiac Function and Angiogenesis in Diabetic Mice

doi: 10.2337/db15-1563

Figure Lengend Snippet: Altered exosomal contents by overexpression of Hsp20 in cardiomyocytes could be delivered to endothelial cells and native cardiomyocytes. Representative Western blots (A) and their quantitative results (B) showed that higher levels of Hsp20, survivin, and p-Akt encased in TG-Exo than WT-Exo. C: SOD1 levels were increased in TG-Exo, compared with WT-Exo (n = 4 for A–C). *P < 0.05 vs. WT-Exo. D: Green dye–labeled exosomes were detectable in the cytosol of MCECs after exposure to green dye PKH67-labeled WT- and TG-Exo, respectively. E and F: Exosomal Hsp20, p-Akt, and survivin were effectively transported to endothelial cells, compared with control conditions (n = 4). *P < 0.05 vs. control conditions; #P < 0.05 vs. WT-Exo–treated samples. β-Actin was used as a loading control. EC, endothelial cell.

Article Snippet: Cardiac Endothelial Cell Proliferation, Migration, and Tube Formation Assays A mouse cardiac endothelial cell (MCEC) line was purchased from CELLutions Biosystems.

Techniques: Over Expression, Western Blot, Labeling, Control

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Coronary Microvascular Dysfunction Is Associated With Augmented Lysosomal Signaling in Hypercholesterolemic Mice

doi: 10.1161/JAHA.124.037460

Figure Lengend Snippet: MCECs were cultured in low glucose DMEM with 5% FBS and treated with 7‐ketocholesterol for the indicated times. A and D , Representative immunofluorescence images and quantification show mitochondrial superoxide levels. Representative immunofluorescence images and quantification show the expression of proinflammatory proteins VCAM‐1 ( B and E ) and CCL2 ( C and F ). G , Representative immunofluorescence images and quantification show the nuclear TFEB positive percentage. Nuclei were stained with DAPI. H , Representative immunoblots and summarized data show the effects of 7‐ketocholesterol on the protein expression levels of microtubule‐associated proteins light chain 3‐II. I , Real‐time reverse transcription polymerase chain reaction analyses of TFEB, LAMP‐1, LAMP‐2A, beclin‐1, microtubule‐associated proteins light chain 3, and p62/SQSTM1 mRNA levels after treatment with 0 or 40 μM 7‐ketocholesterol for 24 hour. Scale bar=20 μm. * vs 0, P <0.05 (n=4–5). CCL2 indicates C‐C motif chemokine ligand 2; LAMP‐1, lysosomal‐associated membrane protein; MCECs, mouse cardiac endothelial cells; TFEB, transcriptional factor EB; and VCAM‐1, vascular cell adhesion molecule 1.

Article Snippet: MCECs (Cedarlane, CLU510) were cultured in low glucose DMEM with 5% FBS, 1% penicillin/streptomycin, and 1 mmol/L of HEPES at 37 °C with 5% CO 2 .

Techniques: Cell Culture, Immunofluorescence, Expressing, Staining, Western Blot, Reverse Transcription, Polymerase Chain Reaction, Membrane

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Coronary Microvascular Dysfunction Is Associated With Augmented Lysosomal Signaling in Hypercholesterolemic Mice

doi: 10.1161/JAHA.124.037460

Figure Lengend Snippet: MCECs were cultured and treated in low glucose DMEM with 5% FBS, pretreated with or without 50 nM BAF for 1 hour, and then cotreated with or without 40 μM of 7‐ketocholesterol for 6 hour. A , Representative immunofluorescence images and quantification show the nuclear TFEB‐positive percentage. Nuclei were stained with DAPI. B through G , MCECs are treated in low‐glucose DMEM with 1% FBS for 2 hour before pretreatment with or without 50 nM of BAF for 1 hour, and then the cells are cotreated with or without 40 μM of 7‐ketocholesterol for 24 hour. B , Representative images of VCAM‐1 and summarized data. C through G , Representative images of FLICA/PI staining and summarized data. H , Cell numbers were detected by using CCK8 kit. Scale bar=20 μm. * vs 0, # vs BAF or 7‐ketocholesterol, P <0.05 (n=4). BAF indicates bafilomycin A1; CCK8, Cell‐Counting Kit 8; MCECs, mouse cardiac endothelial cells; PI, propidium iodide; TFEB, transcriptional factor EB; and VCAM‐1, vascular cell adhesion molecule 1.

Article Snippet: MCECs (Cedarlane, CLU510) were cultured in low glucose DMEM with 5% FBS, 1% penicillin/streptomycin, and 1 mmol/L of HEPES at 37 °C with 5% CO 2 .

Techniques: Cell Culture, Immunofluorescence, Staining, Cell Counting

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Coronary Microvascular Dysfunction Is Associated With Augmented Lysosomal Signaling in Hypercholesterolemic Mice

doi: 10.1161/JAHA.124.037460

Figure Lengend Snippet: MCECs were cultured in low glucose DMEM with 5% FBS, then treated with EZE with or without 7‐ketocholesterol for the indicated time. A , Representative immunofluorescence images and quantification show the effect of ezetimibe on nuclear TFEB positive percentage. B , Representative immunofluorescence images and quantification show the effect of ezetimibe and 7‐ketocholesterol on nuclear TFEB positive percentage. Representative immunofluorescence images and quantification of mitochondrial superoxide ( C ), proinflammatory proteins vascular cell adhesion molecule 1 (VCAM‐1) ( D ) and CCL2 ( E ), and monocyte adhesion ( F ). Scale bar=20 μm. * vs 0, # vs 7‐keto, P <0.05 (n=4). CCL2 indicates C‐C motif chemokine ligand 2; EZE, ezetimibe; MCECs, mouse cardiac endothelial cells; TFEB, transcriptional factor EB; and VCAM‐1, vascular cell adhesion molecule 1.

Article Snippet: MCECs (Cedarlane, CLU510) were cultured in low glucose DMEM with 5% FBS, 1% penicillin/streptomycin, and 1 mmol/L of HEPES at 37 °C with 5% CO 2 .

Techniques: Cell Culture, Immunofluorescence

Journal: Contrast Media & Molecular Imaging

Article Title: Integrated Analysis of lncRNA-miRNA-mRNA ceRNA Network in Mixed Dry Eye Disease

doi: 10.1155/2022/1534142

Figure Lengend Snippet: Verification of ceRNA relation pairs: (a) the expression of miR-671-5p and miR-1934-5p in mouse cornea; (b) the expression of NONMMUT054540.2, miR-1934-5p, and Grm-2 in hypertonic MCECs compared with control group; and (c) the expression of NONMMUT047964.2, miR-671-5p, and Egr-1 in hypertonic MCECs compared with control group.

Article Snippet: The mouse corneal epithelial cell lines (MCECs) are purchased from ICell Bioscience Inc. (Shanghai, China).

Techniques: Expressing, Control

Journal: Contrast Media & Molecular Imaging

Article Title: Integrated Analysis of lncRNA-miRNA-mRNA ceRNA Network in Mixed Dry Eye Disease

doi: 10.1155/2022/1534142

Figure Lengend Snippet: Validation of lncRNA NONMMUT047964.2, regulating the inflammation and apoptosis in vivo: (a) flow cytometric analysis of MCECs apoptosis after treatment; (b) barplot of apoptosis after treatment; (c) the expression of IL-1 β ; (d) the expression of IL-6; (e) the expression of NONMMUT047964.2; (f) the expression of miR-671-5p; and (g) the expression of Egr-1 in MCECs following NaCl treatment.

Article Snippet: The mouse corneal epithelial cell lines (MCECs) are purchased from ICell Bioscience Inc. (Shanghai, China).

Techniques: Biomarker Discovery, In Vivo, Expressing