Journal: Journal of Biomedical Optics

Article Title: Functionalized erythrocyte-derived optical nanoparticles to target ephrin-B2 ligands

doi: 10.1117/1.JBO.24.8.085002

Figure Lengend Snippet: Ephrin-B2 Western blot expression. Western blot showing the expression of ephrin-B2 in hDMVECs infected with an ephrin-B2 lentivirus compared to the ephrin-B2 expression level of hDMVECs not infected with the ephrin-B2 lentivirus. Beta-actin levels are shown as the internal loading control.

Article Snippet: Human dermal microvascular endothelial cells (hDMVECs) were cultured in endothelial cell basal medium (ECBM) with growth supplement (Cell Applications, Inc.) as previously described.

Techniques: Western Blot, Expressing, Infection, Control

Journal: Journal of Biomedical Optics

Article Title: Functionalized erythrocyte-derived optical nanoparticles to target ephrin-B2 ligands

doi: 10.1117/1.JBO.24.8.085002

Figure Lengend Snippet: Cellular fluorescence of control and ephrin-B2 hDMVECs incubated with F-NETs at various ρ * values. (a) Fluorescent images of control and ephbin-B2 hDMVECs after 40 min of incubation at 4°C with F-NETs solutions. All images are falsely colored with the blue and red corresponding to DAPI and ICG NIR emission from the NETs, respectively. Scale bars are 30 μ m . (b) Averaged fluorescence intensity ( I ¯ ) [see Eq. (3)] as a function of ρ * . Cells from 3 to 4 images were analyzed, resulting in 16 to 41 measurements for each combination of nanoparticles and cells. Statistical significance of p < 0.001 is denoted by ***. Only statistically significant populations with the same ρ * value of F-NETs are indicated. (c) Sigmoidal fit to the I ¯ values of the ephrin-B2 hDMVECs versus the ρ * value of F-NETs. Error bars in (b) and (c) represent SDs.

Article Snippet: Human dermal microvascular endothelial cells (hDMVECs) were cultured in endothelial cell basal medium (ECBM) with growth supplement (Cell Applications, Inc.) as previously described.

Techniques: Fluorescence, Control, Incubation

Journal: Cell Communication and Signaling : CCS

Article Title: ER stress inhibition enhances formation of triacylglcerols and protects endothelial cells from lipotoxicity

doi: 10.1186/s12964-024-01682-y

Figure Lengend Snippet: ER stress inhibition restores palmitate-dependent impairment of autophagy and induction of UPR in macro- and microvascular endothelial cells. ( a ) Immunofluorescence of the ER morphology in HUVEC treated with 150 μm of oleate (OA) or palmitate (PA) in combination with DMSO or 2.5 mM 4-PBA. Prior to treatment the cells were transfected with the ER-scarlet probe. Lipid droplets were stained using BODIPY 493/503. The samples were imaged using Zeiss AxioObserver. Scale bar represents 15 μm. (b) Immunoblot analysis of HUVEC treated for 16 h with 150 μm PA in combination with DMSO or 2.5 mM 4-PBA. Cells were lysed in 2x sample buffer and analysed for expression of LC3B, p62, CHOP and Grp78. GAPDH immunblot was used as a loading control. (c) Quantification of immunoblot results from (b). 20 images per condition were analyzed in total in 3 independent experiments. (d) Immunoflourescence analysis of CHOP and p62 expression in HUVEC. The cells were treated as in ( b ), fixed and stained with anti-CHOP and anti-p62 antibodies and DAPI. Scale bar represents 50 μm. (e) Quantification of immunofluorescence analysis depicted in ( d ). For CHOP quantification percentage of CHOP positive cells is displayed and for p62 mean fluorescence intensity (MFI) normalized to control sample. 5 images per condition were analyzed in each of 3 independent experiments. (f) Immunofluorescence analysis of CHOP and p62 expression in HMVEC. Sample preparation and imaging was performed as in ( d ). (g) Quantification of CHOP and p62 staining in HMVEC depicted in ( f ) was performed as described for HUVEC in ( e ). ( i ) FITC labelled dextran leakage assay was performed in HUVEC. Cells were seeded on Transwell inserts and treated for 16 h with BSA + DMSO as a control, 150 µM PA + DMSO or 150 µM PA + 2.5 mM 4-PBA, 150 µM OA or 10 ng/ml TNFa as positive control. The leakage of FITC-labelled 75 kDa Dextran into lower compartment was measured by reading of mean fluorescence intensity at 488 nm using the microplate reader. 3 technical replicates were measured for each condition in 3 independent experiments. GraphPad Prism software and one-way ANOVA with Tukey’s multiple comparison test were used for statistical analysis. Error bars represent standard deviation (SD)

Article Snippet: Human Lung Microvascular Endothelial Cells (HMVEC) were obtained from Sciencell Research Laboratories (#sc-3000) and cultured in Endothelial Cell Medium (ECM)(#sc-1001).

Techniques: Inhibition, Immunofluorescence, Transfection, Staining, Western Blot, Expressing, Control, Fluorescence, Sample Prep, Imaging, Positive Control, Software, Comparison, Standard Deviation