huvec specific medium  (Procell Inc)

Journal: Nature Communications

Article Title: Hexokinase 2-mediated histone H3K18la promotes PAI-1-dependent thrombosis in acute myeloid leukemia via tumor-endothelial crosstalk

doi: 10.1038/s41467-025-65259-0

Figure Lengend Snippet: a PAI-1 in plasma from AML patients ( n = 39) was measured using a commercial assay kit, followed by Pearson correlation analysis with corresponding plasma lactate. b Immunofluorescence staining was performed to evaluate the expression of H3K18la, and PAI-1 in the IVC tissues of thrombus-forming segment. (CD31, an endothelial cell marker; scale bar=50 μm; n = 6 mice per group). c Conditioned media (CM) from NB4 and MOLM13 cells cultured for 12 or 24 hours were co-cultured with HUVECs. Pan-Kla and H3K18la levels in HUVECs were analyzed by Western blot ( n = 3 biological replicates). d, e mRNA and protein expression of PAI-1 in HUVECs co-cultured with conditioned media from AML cell lines were assessed by qPCR and Western blot, respectively ( n = 3 biological replicates). f Plasmin generation in HUVECs co-cultured with conditioned media from AML cell lines was continuously monitored at 405 nm ( n = 3 biological replicates). g Fibrinolysis rate was monitored at 405 nm after co-culture of HUVECs with conditioned medium from AML cell lines, dashed line represents 50% lysis of the clot ( n = 3 biological replicates). h, i HUVECs were transfected with siRNA targeting MCT1 (si MCT1 ), and MCT1 knockdown efficiency was assessed by qPCR and Western blot ( n = 3 biological replicates). j si MCT1 HUVECs were co-cultured with conditioned media from THP-1 cells, and Pan-Kla and H3K18la levels were analyzed by Western blot ( n = 3 biological replicates). k , l si MCT1 HUVECs were co-cultured with conditioned media from THP-1 cells, and PAI-1 mRNA and protein expression were assessed by qPCR and Western blot, respectively ( n = 3 biological replicates). m Plasmin generation was continuously monitored at 405 nm ( n = 3 biological replicates). n Fibrinolysis rate was continuously monitored at 405 nm after co-culture of THP-1-conditioned medium with HUVECs, dashed line represents 50% lysis of the clot ( n = 3 biological replicates). siNC, negative control. Data were analyzed by Pearson correlation analysis ( a ), two-tailed Student’s t test ( d , h , k ), and two-way ANOVA with Sidak’s multiple comparisons test ( f , g , m , n ). Data are presented as mean ± SD. Source data are provided as a Source Data file.

Article Snippet: Human umbilical vein endothelial cells (HUVEC) were cultured in HUVEC cell-specific medium (Procell) and used within 5 passages.

Techniques: Clinical Proteomics, Immunofluorescence, Staining, Expressing, Marker, Cell Culture, Western Blot, Co-Culture Assay, Lysis, Transfection, Knockdown, Negative Control, Two Tailed Test

Journal: Nature Communications

Article Title: Hexokinase 2-mediated histone H3K18la promotes PAI-1-dependent thrombosis in acute myeloid leukemia via tumor-endothelial crosstalk

doi: 10.1038/s41467-025-65259-0

Figure Lengend Snippet: a Schematic diagram: NB4 cells were cultured in [1,2-¹³C] glucose medium, and the labeled supernatant was co-cultured with HUVECs (by figdraw.com). Isotopically labeled metabolites in HUVECs were analyzed by metabolic flux analysis. b Heatmap showing the total levels of metabolites in HUVECs. c Metabolic flux analysis of (M + 2) lactate intensity in HUVECs co-cultured with conditioned media (CM) under different conditions. d Western blot analysis of Pan-Kla and H3K18la in HUVECs co-cultured with conditioned media from HK2 -knockdown NB4 or MOLM13 cells. e , f mRNA and protein levels of PAI-1 in HUVECs co-cultured with conditioned media from AML cells were assessed by qPCR and Western blot, respectively. g Plasmin generation in HUVECs co-cultured with conditioned media from AML cells was monitored at 405 nm. h Fibrinolysis rate in HUVECs co-cultured with conditioned media from HK2 -knockdown AML cells was monitored at 405 nm, dashed line represents 50% lysis of the clot. i Immunofluorescence staining was performed to evaluate the expression of H3K18la, and PAI-1 in the IVC tissues of thrombus-forming segment. (CD31, an endothelial cell marker; n = 6 mice per group; scale bar=50 μm). j ChIP-qPCR analysis of H3K18la enrichment at the PAI-1 promoter in HUVECs co-cultured with conditioned media from HK2 -knockdown NB4 cells. b – h , j n = 3 biological replicates. Data were analyzed by two-tailed Student’s t test ( c , e , j ), two-way ANOVA with Sidak’s multiple comparisons test ( g , h ). Data are presented as mean ± SD. Source data are provided as a Source Data file.

Article Snippet: Human umbilical vein endothelial cells (HUVEC) were cultured in HUVEC cell-specific medium (Procell) and used within 5 passages.

Techniques: Cell Culture, Labeling, Western Blot, Knockdown, Lysis, Immunofluorescence, Staining, Expressing, Marker, ChIP-qPCR, Two Tailed Test

Journal: BMC Cancer

Article Title: Exploration of RNA-binding proteins identified RPS27 as a potential regulator associated with Kaposi’s sarcoma development

doi: 10.1186/s12885-025-13790-0

Figure Lengend Snippet: siRPS27 increased the pro-tumor signatures of HUVECs. A Bar plot showing the expression level of RPS27 mRNA in WT, siNC, and siRPS27 samples. B Western blot results show the expression level of RPS27 protein in three groups. The right panel shows the quantitative result. C Cell cycle results show the percentages of three cellular stages in three groups. The right panel shows the quantitative result. D Bar plot showing the cellular proliferation results in three groups. E Cell migration results show the migrated cell numbers in three groups. The right panel shows the quantitative result. F Cell invasion results show the invaded cell numbers in three groups. The right panel shows the quantitative result. G Tube formation results show the tube length in three groups. The right panel shows the quantitative result. * p -value < 0.05; ** p -value < 0.01; *** p -value < 0.001; Student’s t -test

Article Snippet: After culturing each group of cells in serum-free HUVEC cell-specific culture medium for 48 h, a cell suspension was prepared, and 50 μL of cell suspension (1 × 10 5 cells) was seeded in the plate coated with Matrigel and incubated for another 12 h. Representative photographs were subsequently captured under the microscope (IX53, OLYMPUS, Japan) after co-culture.

Techniques: Expressing, Western Blot, Migration