Journal: Scientific Reports

Article Title: Networked lymphatic endothelial cells in a transplanted cell sheet contribute to form functional lymphatic vessels

doi: 10.1038/s41598-022-26041-0

Figure Lengend Snippet: Secretion of growth factors by adipose-derived stem cells (ASCs) and the formation of networks by co-culture with endothelial cells (ECs) or lymphatic endothelial cells (LECs). ( a ) The levels of vascular endothelial growth factor-C (VEGF-C), VEGF-A, hepatocyte growth factor (HGF) and basic fibroblast growth factor (bFGF) in the medium bathing cultured ASCs were measured using enzyme-linked immunosorbent assays. HGF and VEGF-A were both secreted by ASCs, whereas the levels of secreted VEGF-C and b-FGF were much lower. ( b ) Green fluorescent protein (GFP)-expressing ECs and ASCs co-cultured at a ratio of 1:4. Networks of vascular ECs were observed, and the edges of the vascular EC network were sharp (arrows). ( c ) GFP-LECs and ASCs co-cultured at a ratio of 1:4. The LECs formed networks, but in contrast to vascular ECs, rounded/bulging structures were observed at the edges of the LEC network (arrowheads). ( d ) Merge image of ( c ) and phase-contrast image. ASCs shows random pattern with no specific character which do not form own network or support LEC network directly.

Article Snippet: The concentrations of VEGF-C, VEGF-A, HGF, and bFGF were measured by enzyme-linked immunosorbent assay (ELISA) using a Human VEGF-C ELISA Kit (P49767, RayBiotech, USA), LBIS Human VEGF ELISA Kit (631-40831, Fujifilm Wako, Japan), AuthentiKine Human HGF ELISA Kit (KE00168, Proteintech, USA) and Human bFGF ELISA Kit (P09038, RayBiotech, USA) in accordance with the manufacturers’ instructions.

Techniques: Derivative Assay, Co-Culture Assay, Cell Culture, Expressing

Journal: International Journal of Biological Sciences

Article Title: N 6 -methyladenosine Reader IGF2BP2-modified HMMR Promotes Non-small Cell Lung Cancer Metastasis via Interaction with MAP4K4

doi: 10.7150/ijbs.104097

Figure Lengend Snippet: MAP4K4 inhibitor GNE-495 reduces NSCLC cell metastasis in vivo . (A) Flowchart illustrating the NSCLC cell in vivo metastasis model. (B) Relative mRNA and protein expression levels of HMMR and MMP1 in the A549 cells used to establish the in vivo metastasis model. (C) Photographs showing lung metastatic nodules in mice following injection of A549 cells overexpressing HMMR or the control vector, with treatment by DMSO or GNE-495. Metastatic nodules are indicated by red arrowheads. (D) Quantification of lung metastatic nodules across the four experimental groups. (E) The lung weights of the mice in each group after execution. (F) Hematoxylin and eosin (H&E) staining of lung tissue to assess micrometastatic foci. Representative histological images of micrometastatic foci in the four groups. Micrometastatic foci are indicated by blue arrowheads.

Article Snippet: From the fifth week post-injection, mice received daily intraperitoneal injections of either DMSO (3 mg/kg) or the MAP4K4 inhibitor GNE-495 (HY-100343, MCE, China) (3 mg/kg).

Techniques: In Vivo, Expressing, Injection, Control, Plasmid Preparation, Staining

Journal: Cancers

Article Title: Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation

doi: 10.3390/cancers13081845

Figure Lengend Snippet: Flow cytometry antibodies used.

Article Snippet: CD3-VB , Miltenyi , 130-133-133 , BW264/56 , .

Techniques: Flow Cytometry, In Vivo, In Vitro

Journal: Cancers

Article Title: Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation

doi: 10.3390/cancers13081845

Figure Lengend Snippet: Murlentamab opsonization of SKOV3-R2 + activates an effective anti-tumor T cell immune response. SKOV3-R2 + ovarian tumor cells were labeled with different 3C23K antibodies (3C23K-FcKO control, 3C23K-CHO normally fucosylated or murlentamab the low fucosylated form) and cultured in the presence of human monocyte-derived macrophages from healthy donors unstimulated (M0) or stimulated with M-CSF and IL-10 (TAMs). After 3 days of co-culture, activated T cells coming from the same healthy donor were added in the culture well for 4 more days. ( A ) The CD4 + Th1/Th2 polarization profile, ( B ) the proportion of CD3 + CD4 + CD25 + regulatory T cells and ( C ) the activation of T CD8 + cells were determined by flow cytometry after four days of co-culture. Data shown (boxplots) are the results from two different experiments (performed with two different healthy donors). * p < 0.05; ** p < 0.01; *** p < 0.001. p values were determined using one-way ANOVA analysis followed by Tukey’s multiple comparisons test.

Article Snippet: CD3-VB , Miltenyi , 130-133-133 , BW264/56 , .

Techniques: Labeling, Cell Culture, Derivative Assay, Co-Culture Assay, Activation Assay, Flow Cytometry

Journal: Epigenetics

Article Title: Epigenetic aberrations of gene expression in a rat model of hepatocellular carcinoma

doi: 10.1080/15592294.2022.2069386

Figure Lengend Snippet: Overexpression and DNA hypomethylation of MMP12 in human HepG2 HCC cells is accompanied by TET1 increased occupancy at MMP12 promoter region . (a-b) Expression of MMP12 in human HepG2 HCC cells and in human primary hepatocytes (NorHep), as measured by qRT-PCR (a) and western blot (b). In western blot analyses, reactive bands were visualized using the chemiluminescent protocol in the ChemiDoc MP Imaging System (Bio-Rad) and analysed using the Image Lab software. Protein loading was normalized with respect to total sample protein loaded using free stain gels. (c) DNA methylation levels across three CpG sites within MMP12 promoter region in HepG2 and NorHep, as measured by pyrosequencing. (d) TET1 binding within MMP12 promoter region in HepG2 and NorHep, as measured by quantitative chromatin immunoprecipitation (qChIP). (e) Number of HepG2 and NorHep cells that invaded through extracellular matrix within 24 hr, as assessed by Boyden chamber invasion assay. Results are expressed as mean ± SD; n = 3, *** P < 0.001.

Article Snippet: Human HCC HepG2 cells were purchased from ATCC (HB8065) and human untransformed hepatocytes (normal hepatocytes, NorHep) were obtained from Celprogen (33,003–02).

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Western Blot, Imaging, Software, Staining, DNA Methylation Assay, Binding Assay, Chromatin Immunoprecipitation, Invasion Assay