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STDP <t>attenuates</t> <t>vascular</t> 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 <t>MVECs</t> 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
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Thermo Fisher mvec monolayers
STDP <t>attenuates</t> <t>vascular</t> 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 <t>MVECs</t> 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
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ATCC microvascular endothelial cells
STDP <t>attenuates</t> <t>vascular</t> 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 <t>MVECs</t> 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
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PromoCell human lung microvascular endothelial cells hlmec
STDP <t>attenuates</t> <t>vascular</t> 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 <t>MVECs</t> 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
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PromoCell human cardiac microvascular endothelial cells
STDP <t>attenuates</t> <t>vascular</t> 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 <t>MVECs</t> 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
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ATCC human microvascular endothelial cells
STDP <t>attenuates</t> <t>vascular</t> 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 <t>MVECs</t> 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
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PromoCell primary human cardiac microvascular endothelial cells
STDP <t>attenuates</t> <t>vascular</t> 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 <t>MVECs</t> 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
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ATCC human microvascular endothelial cells humec
The role of endogenous IFITMs in NiV and HeV pseudovirus entry into human <t>endothelial</t> and epithelial cells. The endogenous IFITMs mRNA levels in <t>HuMEC</t> ( 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).
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PromoCell hcmecs
The role of endogenous IFITMs in NiV and HeV pseudovirus entry into human <t>endothelial</t> and epithelial cells. The endogenous IFITMs mRNA levels in <t>HuMEC</t> ( 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).
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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

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

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

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

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).

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