Journal: Molecular Vision

Article Title: Wnt3a promotes epithelial–mesenchymal transition, migration, and proliferation of lens epithelial cells

doi:

Figure Lengend Snippet: Wnt3a induces the EMT of HLE B-3 cells. A : Wnt3a-overexpressing cells had an irregular shape, whereas control cells had a spindle-shaped morphology (original magnification 400×). B : western blot analysis detected down-regulated epithelial protein E-cadherin in Wnt3a-overexpressing HLE B-3 cells compared with control cells. In contrast, the expression of mesenchymal protein fibronectin was upregulated in Wnt3a-overexpressing cells. C : Immunocytofluorescence showed that the expression of E-cadherin protein (green) was down-regulated in Wnt3a-overexpressing cells compared with control cells, whereas that of fibronectin (red) increased. DAPI (blue) was used for nuclear staining. Merged images are shown in the right panel.

Article Snippet: The immortalized human lens epithelial (HLE) cell line HLE B-3 was obtained from American Type Culture Collection (Manassas, VA).

Techniques: Control, Western Blot, Expressing, Staining

Journal: JCI Insight

Article Title: Whole-body deletion of Endospanin 1 protects from obesity-associated deleterious metabolic alterations

doi: 10.1172/jci.insight.168418

Figure Lengend Snippet: ( A ) Detection of Flag-CD36 in 6Myc-Endo1 immunoprecipitate from HEK293T cells coexpressing 6Myc-Endo1 and Flag-CD36 compared with HEK293T cells transfected with Flag-CD36 alone. Representative blots of 3 independent experiments. ( B ) Detection of 6Myc-Endo1 after Flag-CD36 immunoprecipitation from HEK293T cells coexpressing 6Myc-Endo1 and Flag-CD36 compared with HEK293T cells transfected with 6Myc-Endo1 alone. Representative blots of 3 independent experiments. ( C ) Kinetics of Endo1 and CD36 protein expression in differentiated adipocytes during differentiation of adipocyte precursors isolated from the stromal vascular fraction of the s.c. adipose tissue of WT mice into mature white adipocytes. Results are expressed as mean ± SEM of 8 independent experiments. One representative Western blot is shown. ( D ) Confocal immunofluorescence detection and colocalization of Endo1 (rabbit anti-Endo1) and CD36 (goat anti-CD36) in differentiated adipocytes. Nuclei (blue) are stained with fluorescent DAPI dye. Scale bar: 20 μm. Representative images of 4 independent experiments. ( E ) Detection of endogenous Endo1 and CD36 after immunoprecipitation with Endo1 or CD36 antibodies from lysates of visceral adipose tissue (VAT), gonadal adipose tissue (GAT), and s.c. adipose tissue (SAT) of WT and KO mice. Endo1-KO adipocytes and tissues were used as negative controls. The molecular weights of protein markers are indicated (kDa). Representative blots of 2 independent experiments.

Article Snippet: Next, cells were incubated with primary Endo1 (1:500) (generated as described previously; ref. ) and primary CD36 (1:1,000) (RD Systems, catalog AF2519), primary Endo1 (1:500) and primary 53K (1:1,000, Abcam, catalog ab27043), or primary GM130 (1:1,000, Abcam, catalog ab169276) antibodies overnight at 4°C.

Techniques: Transfection, Immunoprecipitation, Expressing, Isolation, Western Blot, Immunofluorescence, Staining

Journal: JCI Insight

Article Title: Whole-body deletion of Endospanin 1 protects from obesity-associated deleterious metabolic alterations

doi: 10.1172/jci.insight.168418

Figure Lengend Snippet: ( A ) Cell surface expression of CD36 in mature white adipocytes. ** P < 0.01 versus WT. Results are expressed as mean ± SEM ( n = 4). Two-tailed t test. ( B ) Total CD36 expression in gonadal adipose tissue (GAT) and s.c. adipose tissue (SAT) . The molecular weights of protein markers are indicated (kDa). Results are expressed as mean ± SEM ( n = 5). ( C ) Immunofluorescence images of CD36 cell surface expression in differentiated white adipocytes (left panel). Level of cellular fluorescence determined by corrected total cell fluorescence per area (CTCF/area). Results are expressed as mean ± SEM ( n = 9). *** P < 0.005 versus WT. Two-tailed t test (right panel). Scale bar: 20 μm. ( D ) Lipid uptake in differentiated adipocytes, mature adipocytes, and differentiated myotubes. Results are expressed as mean ± SEM ( n = 5–6). * P < 0.05; ** P < 0.01; **** P < 0.001 versus basal. † P < 0.05; †† P < 0.01; †††† P < 0.001 versus WT. One-way ANOVA with Bonferroni correction.

Article Snippet: Next, cells were incubated with primary Endo1 (1:500) (generated as described previously; ref. ) and primary CD36 (1:1,000) (RD Systems, catalog AF2519), primary Endo1 (1:500) and primary 53K (1:1,000, Abcam, catalog ab27043), or primary GM130 (1:1,000, Abcam, catalog ab169276) antibodies overnight at 4°C.

Techniques: Expressing, Two Tailed Test, Immunofluorescence, Fluorescence