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: Cancer Communications

Article Title: NAT10‐mediated ac 4 C‐modified ANKZF1 promotes tumor progression and lymphangiogenesis in clear‐cell renal cell carcinoma by attenuating YWHAE‐driven cytoplasmic retention of YAP1

doi: 10.1002/cac2.12523

Figure Lengend Snippet: NAT10 promoted tumor lymphangiogenesis in ccRCC. (A) GSEA showed the associations between the VEGF signaling and the NAT10 mRNA levels in ccRCC. FDR q < 25% was considered statistically significant. Patients were categorized into low and high subgroups using median expression (50%) as the cut‐off. (B) Cell proliferation curves of HLECs treated with CM from the NAT10‐overexpressed ccRCC cells and the control cells ( n = 4) ( t ‐test for statistics). (C) Cell proliferation curves of HLECs treated with CM from the NAT10‐knockdown ccRCC cells and the control cells ( n = 4) ( t ‐test for statistics). (D) Transwell assays for HLECs treated with CM from the NAT10‐overexpressed ccRCC cells and the control cells ( n = 3) ( t ‐test for statistics). (E) Transwell assays for HLECs treated with CM from the NAT10‐knockdown ccRCC cells and the control cells ( n = 3) ( t ‐test for statistics). (F) Tube formation assays for HLECs treated with CM from the NAT10‐overexpressed ccRCC cells and the control cells ( n = 3) ( t ‐test for statistics). (G) Tube formation assays for HLECs treated with CM from the NAT10‐knockdown ccRCC cells and the control cells ( n = 3) ( t ‐test for statistics). (H) ELISAs were used to detect the VEGFD concentrations in CM from ccRCC cells with NAT10 overexpression or knockdown ( n = 3) ( t ‐test for statistics). (I) IHC of NAT10, VEGFC/D, and LYVE1 in subcutaneous tumors from the NAT10‐knockdown group and the control group ( n = 5) (Mann‐Whitney U for statistics). Results represented at least three independent experiments (* P < 0.05, ** P < 0.01, *** P < 0.001). Abbreviations: NAT10, N‐acetyltransferase 10; ccRCC, clear‐cell renal cell carcinoma; GSEA, gene set enrichment analysis; FDR, false discovery rate; NES, normalized enrichment score; VEGF, vascular endothelial growth factor; HLEC, human lymphatic endothelial cell; CM, conditioned medium; ELISA, Enzyme‐linked immunosorbent assay; VEGFC/D, vascular endothelial growth factor‐C/D; LYVE1, lymphatic vessel endothelial hyaluronan receptor 1; KEGG, kyoto encyclopedia of genes and genomes.

Article Snippet: The VEGFC/D concentrations in CM from ccRCC cells were detected using human VEGFC/D ELISA Kits (#E‐EL‐H1600c and #E‐EL‐H1601c, Elabscience, Wuhan, Hubei, China) following the manufacturer's protocol.

Techniques: Expressing, Control, Knockdown, Over Expression, MANN-WHITNEY, Enzyme-linked Immunosorbent Assay