Journal: International Journal of Molecular Sciences

Article Title: Gene Expression Changes in Cytokine and Chemokine Receptors in Association with Melanoma Liver Metastasis

doi: 10.3390/ijms24108901

Figure Lengend Snippet: The effect of human hepatic sinusoidal endothelial cells conditioned media (HHSEC-CM) on the invasion of melanoma cell lines (WM1366, WM278, WM793B, WM983A, WM1361, and WM3248) compared to the effect of the unconditioned medium as a control. Results are presented as the ratio (mean ± SE, n = 3) of HHSEC-CM-induced and control invaded cells. The asterisk indicates a statistically significant difference between HHSEC-CM induced data and control data (Mann–Whitney test: * p < 0.05). The results of the invasion assay are shown as the mean of three independent experiments.

Article Snippet: Human hepatic sinusoidal endothelial cells (HHSEC) were obtained from ScienCell Research Laboratories, Inc. (Carlsbad, CA, USA) and cultured in a 37 °C, 5% CO 2 incubator according to the recommended protocol.

Techniques: Control, MANN-WHITNEY, Invasion Assay

Journal: International Journal of Molecular Sciences

Article Title: Gene Expression Changes in Cytokine and Chemokine Receptors in Association with Melanoma Liver Metastasis

doi: 10.3390/ijms24108901

Figure Lengend Snippet: Association between chemokine- and cytokine-receptor expression of melanoma cells and the invasive capacity responding to human hepatic sinusoidal endothelial cells conditioned media (HHSEC-CM). ( A ) The Venn diagram illustrates the significant gene expression alterations in cell lines with stable invasiveness ( a ) and in those with increased invasiveness ( b ), indicating the IL11RA gene in the overlapping region of the two sets of gene expression. Arrows indicate overexpression (↑) and downregulation (↓) in the selected invasive cells compared to the non-invasive populations. ( B ) Unsupervised hierarchical clustering of the significantly differentially expressed genes in the melanoma cell lines with increased invasiveness. The separated non-invasive (NON-INV) and invasive (INV) cells are displayed vertically, and the genes are displayed horizontally. The heat map was generated from the seven significantly expressed genes using www.heatmapper.ca accessed on 4 May 2023. ( C ) Relative mRNA expressions of the IL11RA gene in selected non-invasive and invasive cells according to the invasive capacity of the cell lines. The cell lines that exhibited stable invasiveness (left part) showed nearly imperceptible differences, while the cell lines with increased invasiveness (right part) demonstrated a significantly higher level of IL11RA. The data are presented as the mean ± SD of three independent experiments. An asterisk indicates statistically significant differences (Mann–Whitney–Wilcoxon test, * p < 0.05); the qRT-PCR results are presented as the mean ratio of 2 −ΔCt values between invasive and non-invasive populations of three independent experiments.

Article Snippet: Human hepatic sinusoidal endothelial cells (HHSEC) were obtained from ScienCell Research Laboratories, Inc. (Carlsbad, CA, USA) and cultured in a 37 °C, 5% CO 2 incubator according to the recommended protocol.

Techniques: Expressing, Gene Expression, Over Expression, Generated, MANN-WHITNEY, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Gene Expression Changes in Cytokine and Chemokine Receptors in Association with Melanoma Liver Metastasis

doi: 10.3390/ijms24108901

Figure Lengend Snippet: Relative protein expression profiles of human hepatic sinusoidal endothelial cells (HHSECs) co-cultured with different melanoma cell lines. The intensity of the reference is displayed at 100%. Numbers beside the columns indicate the protein expression as a percentage of the intensity of the reference spots on the array. Protein expression in control HHSECs is indicated in green; protein expression in HHSECs co-cultured with melanoma cell lines with stable invasiveness is indicated in blue; and protein expression in HHSECs co-cultured with cell lines with increased invasiveness is indicated in red.

Article Snippet: Human hepatic sinusoidal endothelial cells (HHSEC) were obtained from ScienCell Research Laboratories, Inc. (Carlsbad, CA, USA) and cultured in a 37 °C, 5% CO 2 incubator according to the recommended protocol.

Techniques: Expressing, Cell Culture, Control

Journal: Journal of Leukocyte Biology

Article Title: Frontline Science: HIV infection of Kupffer cells results in an amplified proinflammatory response to LPS

doi: 10.1189/jlb.3HI0516-242R

Figure Lengend Snippet: (A) Immunofluorescence CD68 staining (green) reveals KCs within the hepatic sinusoids (left, 200× original magnification; right, 640× original magnification). (B) Number of KCs/mm2 quantitated and shown graphically. HCV and HIV/HCV, n = 6; normal, n = 3. (C) KC morphology (phase-contrast photomicrograph original magnification, 200×). (D) Representative FACS analyses of CD68 expression directly after KC isolation (gray histogram) and adherence enrichment of KCs (white histogram). (E) FACS demonstrating quantitative expression of CD68 in primary liver-derived KCs directly after isolation, with adherence enrichment, and after HIV infection (n = 10). (F) RT-qPCR confirms low expression of CD3 and CD31 in KC preparations compared with human T cells and liver sinusoidal endothelial cells (LSECs), respectively (n = 3 T cells and LSECs; n = 5 KCs).

Article Snippet: RT-qPCR on RNA extracted from KCs was performed to determine the presence of significant contamination with T cells (primers to CD3) or endothelial cells (primers to CD31). cDNA from primary CD4 + T cells and human hepatic sinusoidal endothelial cells (Cat. #5000; ScienCell, Carlsbad, CA, USA) were used as positive controls for CD3 and CD31 expression, respectively.

Techniques: Immunofluorescence, Staining, Expressing, Isolation, Derivative Assay, Infection, Quantitative RT-PCR