human microvascular endothelial cells humec  (ATCC)

Journal: bioRxiv

Article Title: IFITM1 inhibits Henipavirus membrane fusion by trapping ephrinB2 receptors in fusion-unfavorable membrane nanodomains

doi: 10.64898/2026.05.06.723334

Figure Lengend Snippet: The role of endogenous IFITMs in NiV and HeV pseudovirus entry into human endothelial and epithelial cells. The endogenous IFITMs mRNA levels in HuMEC ( a-c ) and HEK293T cells ( g-i ). Cells were transfected with siRNAs targeting IFITM proteins and scrambled siRNA (NC). IFN-α2b was used to stimulate the expression of IFITM proteins. The mRNA expression levels were detected using qPCR and normalized to that of NC at the untreated condition (-IFN-α). d and j , endogenous IFITM1,2,3 proteins expression in HuMEC ( d ) and HEK293T cells ( j ) upon siRNA knockdown analyzed by Western Blot. IFITMs were detected by anti-IFITM antibodies, and GAPDH was a loading control. The entry of NiV/VSV pp and HeV/VSV pp to HuMEC ( e and f ) and HEK293T cells ( k and l ). NiV and HeV glycoproteins were pseudotyped to VSV particles in which the VSV-G gene was replaced with the Renilla luciferase gene. Virus entry was measured by luminescence intensity and normalized to that of scrambled siRNA at the untreated condition (NC, −IFN-α). Virus entry levels in siRNA-transfected cells were compared with those transfected with scrambled siRNA (NC) under respective −IFN-α and +IFN-α conditions. Virus entry in IFN-α–treated cells (+IFN-α) was further compared with that in resting cells (−IFN-α) and labeled with a bracket. Bars represent means ± SEM. Results from at least 3 independent experiments are shown. p values were obtained using one-way analysis of variance (ANOVA) with post hoc correction (nonsignificant [ns], p > 0.05; *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001).

Article Snippet: Human microvascular endothelial cells (HuMEC) (ATCC CRL-4060) were cultured in Vascular cell basal medium (ATCC, PCS-100-030) with Microvascular endothelial cell growth kit – BBE (ATCC, PCS-110-040) and 0.5 ug/ml puromycin (10 mg/ml stock, Gibco A1138-03).

Techniques: Transfection, Expressing, Knockdown, Western Blot, Control, Luciferase, Virus, Labeling

Journal: Chinese Medicine

Article Title: Shexiang Tongxin Dropping Pills targeting Piezo1/Ca 2+ /NLRP3 axis attenuates vascular endothelial inflammation

doi: 10.1186/s13020-026-01402-3

Figure Lengend Snippet: STDP attenuates vascular endothelial dysfunction by NLRP3 inhibition. A – D The expression levels of NLRP3, pro-IL-1β and cle-IL-1β were analyzed by western blotting and the relative protein expression level were determined by densitometric analysis. E , F The expression levels of ICMA-1 were analyzed by western blotting and the relative protein expression level were determined by densitometric analysis. G, H Representative fluorescent confocal images of VCAM-1 (green) and DAPI (blue) in MVECs and the summarized data of the immunofluorescence intensity, scale bar = 10 μm. N = 4–6, * P < 0.05, *** P < 0.001 compared to control group, # P < 0.05, ## P < 0.01, ### P < 0.001 compared to LCWE group

Article Snippet: MVECs (murine vascular endothelial cells) were purchased from ATCC (CRL-2586, Shanghai, China) and cultured in a thermostat incubator at 37 °C with 5% carbon dioxide concentration using high glucose medium containing 10% fetal bovine serum (Gibico, USA).

Techniques: Inhibition, Expressing, Western Blot, Immunofluorescence, Control

Journal: Chinese Medicine

Article Title: Shexiang Tongxin Dropping Pills targeting Piezo1/Ca 2+ /NLRP3 axis attenuates vascular endothelial inflammation

doi: 10.1186/s13020-026-01402-3

Figure Lengend Snippet: Piezo1 knockdown abrogates STDP-mediated endothelial protection. A , B Knockdown Piezo1 representative images as well as statistical graphs. C , D The protein expression levels of NLRP3 was analyzed by Western blotting and the relative protein expression level were determined by densitometric analysis. E , F Immunofluorescence double staining to co-localize NLRP3 with Caspase1. G, H The levels of IL-18 and IL-1β in cell culture supernatants were measured by ELISA. I, J The expression levels of pro-Caspase-1, cle-Caspase-1 were analyzed by Western blotting and the relative protein expression level were determined by densitometric analysis. K, L The protein expression levels of ICAM-1 was analyzed by Western blotting and the relative protein expression level were determined by densitometric analysis. M , N Representative fluorescent confocal images of VCAM-1 (green) and DAPI (blue) in MVECs and the summarized data of the immunofluorescence intensity, scale bar = 10 μm. N = 4–6, * P < 0.05, ** P < 0.01, *** P < 0.001 compared to control group, # P < 0.05, ## P < 0.01, ### P < 0.001 compared to LCWE group

Article Snippet: MVECs (murine vascular endothelial cells) were purchased from ATCC (CRL-2586, Shanghai, China) and cultured in a thermostat incubator at 37 °C with 5% carbon dioxide concentration using high glucose medium containing 10% fetal bovine serum (Gibico, USA).

Techniques: Knockdown, Expressing, Western Blot, Immunofluorescence, Double Staining, Cell Culture, Enzyme-linked Immunosorbent Assay, Control

Journal: bioRxiv

Article Title: Temporal AI model predicts drivers of cell state trajectories across human aging

doi: 10.64898/2026.03.30.715396

Figure Lengend Snippet: ( A ) Predicted impact of in silico inhibition vs. slope of expression change across human aging for genes in cardiac fibroblasts. ( B ) Concordant genes whose inhibition was predicted to be rejuvenating were upregulated across aging, and vice versa. ( C ) Predicted impact of in silico inhibition of genes across the indicated cardiac cell types. ( D ) Beta-gal staining and cell counts quantifying senescence of primary human cardiac capillary endothelial cells in response to predicted pro-aging perturbation of ZBTB16 inhibition. *p<0.05, Wilcoxon rank sums, n=4. ( E ) Predicted impact of in silico perturbation and expression change across human aging and in telomere-shortened mice in each cardiac cell type for top predicted cardiomyocyte age-modulating targets. ( F ) PCA plot and ( G ) differential gene expression heatmap of transcriptional response (measured by bulk RNA sequencing) to predicted pro-aging perturbations (AAV overexpression of indicated genes vs. GFP) in human iPSC-derived cardiomyocytes. p<0.05, Wald test with BH correction, n=4. ( H ) Gene set enrichment of genes differentially expressed in response to all predicted pro-aging perturbations in human iPSC-derived cardiomyocytes (hypergeometric test with g:Set Counts and Sizes (g:SCS) correction). ( I ) Slowed calcium cycle kinetics and ( J ) rhythm irregularities in response to predicted pro-aging perturbations (AAV overexpression of indicated genes vs. GFP) in human iPSC-derived cardiomyocytes. *p<0.05, Wilcoxon rank sums with BH correction. Rhythm n=10, time to peak n=180, 111, 139, 37, 237, 150, 191 (bars left to right), decay n=169, 30, 86, 11, 213, 105, 172 (bars left to right). ( K ) Differential gene expression heatmap of transcriptional response (measured by bulk RNA sequencing) to predicted pro-aging perturbations (AAV overexpression of indicated genes vs. GFP) in human iPSC-derived cardiomyocytes or human primary cardiac fibroblasts where dysregulation of the given gene was statistically significant across all four conditions. Statistical significance of differential expression: p<0.05, Wald test with BH correction, n=4. Statistical significance of overlap of shared dysregulation between the four conditions: p<0.05, one-sided binomial test. RASG.=RASGEF1B. ( L ) Beta-gal staining quantifying senescence of primary human cardiac fibroblasts in response to predicted pro-aging perturbations (AAV overexpression of indicated genes). *p<0.05, Wilcoxon rank sums with BH correction, empty n=6, P4HA1 and RASGEF1B n=4. ( M ) Schematic of in vivo validation experiment. o/e=overexpression. ( N ) In vivo systolic function echocardiographic measurements at 6 weeks post-AAV9 injection. EF: GFP n=8, P4ha1 n=10, Rasgef1b n=10; GLS: GFP n=8, P4ha1 n=9, Rasgef1b n=8. *p<0.05 one-way ANOVA with multiple hypothesis correction.

Article Snippet: Human cardiac capillary endothelial cells were purchased from PromoCell (C-12285) and cultured in endothelial basal medium (EBM, CC-3121, Lonza) supplemented with 10% fetal bovine serum (10100147, Gibco).

Techniques: In Silico, Inhibition, Expressing, Staining, Gene Expression, RNA Sequencing, Over Expression, Derivative Assay, Quantitative Proteomics, In Vivo, Biomarker Discovery, Injection