Journal: International Journal of Biological Sciences

Article Title: CD137L-macrophage induce lymphatic endothelial cells autophagy to promote lymphangiogenesis in renal fibrosis

doi: 10.7150/ijbs.66781

Figure Lengend Snippet: CD137L promotes the proliferation, migration and tube formation of lymphatic endothelial cells. (A) The proliferation of LECs treated with CD137L or CD137L and inhibitory for 24 h, 48 h, and 72 h was examined by CCK-8 assay. The migration of LECs was measured with the cell scratch test (B) and Transwell migration assay (D) after CD137L or CD137L and inhibitory treatment for 12 h. The histograms represent migrated (C, E) cells per field. (F) A tube formation assay was used to test the lymphangiogenesis capacity of LECs after CD137L or CD137L and inhibitory treatment for 3 h (image magnification, 100×). The histograms represent the branch number per field. (G, H, I). The error bars represent the SEM. *P< 0.05, **P < 0.01; ***P < 0.001. The data were pooled from three independent experiments.

Article Snippet: The mouse lymph node endothelial cell line (SVEC4-10, LECS) was purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Migration, CCK-8 Assay, Transwell Migration Assay, Tube Formation Assay

Journal: International Journal of Biological Sciences

Article Title: CD137L-macrophage induce lymphatic endothelial cells autophagy to promote lymphangiogenesis in renal fibrosis

doi: 10.7150/ijbs.66781

Figure Lengend Snippet: CD137L induced autophagy of lymphatic endothelial cells via mTOR pathway. (A) Representative electron micrographs of CD137L-induced autophagosome in SVEC4-10 cells that were either stimulated with inhibitory, or with 3-MA for 24 h. (B) The protein expression levels of autophagy-associated proteins (Atg5, Atg7 and Atg12) and autophagic substrates (LC3 and p62) were analyzed in SVEC4-10 cells after CD137L, inhibitory or 3-MA treatment for 24 h by Western blot. (C) SVEC4-10 cells were infected with RFP-GFP-LC3-expressing lentivirus. Cells were untreated or treated with CD137L, inhibitory or 3-MA for 24 h. Fluorescence was examined by confocal microscopy. Transwell migration assay (D), scratch test (E) and tube formation assay (F) stimulated with CD137L for 12 h, and with or without 3-MA treatment. (G) The protein expression levels of phosphorylated PI3K, Akt and mTOR were analyzed in SVEC4-10 cells after CD137L or inhibitory or 3-MA treatment for 24 h by Western blot. (H) SVEC4-10 cells were knocked down Atg5 and Atg7 by using siRNAs, and stimulated with CD137L for 24h, the protein expression levels of autophagy-associated proteins (Atg5, Atg7 and Atg12) and autophagic substrates (LC3 and p62) were detected by Western blotting.

Article Snippet: The mouse lymph node endothelial cell line (SVEC4-10, LECS) was purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Expressing, Western Blot, Infection, Fluorescence, Confocal Microscopy, Transwell Migration Assay, Tube Formation Assay

Journal: Nature Communications

Article Title: Hepatocytes functionally reprogrammed by KIAA1199-high colorectal cancer cells favour the accumulation of pro-metastatic Egr1 + neutrophils

doi: 10.1038/s41467-026-69250-1

Figure Lengend Snippet: A Subcutaneous tumor growth in mice co-injected with CRC cells and either Egr1⁺ or control neutrophils. Tumor volume was monitored for 21 days. Inset shows representative tumors at endpoint ( n = 5 mice, two-way ANOVA). B CD31 immunofluorescence (green) and vascular density quantification in subcutaneous tumors. Yellow arrowheads indicate endothelial structures ( n = 5 mice, Student’s t test). Scale bars: 100 µm. C mIHC of pre-metastatic livers (day 14) showing spatial association of Egr1 + neutrophils (white arrowheads) and CD31⁺ vessels (yellow arrowheads). Scale bars: 100 µm. D , E Co-localization analysis of Egr1⁺ neutrophils and CD31⁺ vasculature in mouse ( D , n = 2 mice) and human ( E , n = 2 patients) CRLM samples, highlighting the tumor boundary. Scale bars: 100 µm. F , I SVEC4-10 endothelial cell viability (CCK-8) ( n = 3 independent experiments, Student’s t test). G , J Endothelial functional assays using supernatants from triple co-cultures treated with Rog or WRW4 ( n = 3 independent experiments, Student’s t test). H , K Endothelial migration (scratch wound assay) ( n = 3 independent experiments, Student’s t test). L Representative tube formation images (HUVEC and SVEC4-10) treated with neutrophil-conditioned media ( n = 3 independent experiments). Scale bars: 20 µm. M , N Quantification of vessel numbers ( M ) and junction points ( N ) ( n = 3 independent experiments, Student’s t test). Where applicable, all statistical tests are two-sided. Data are presented as mean ± s.d. Source data and exact p values are provided as a Source Data file.

Article Snippet: Murine colorectal cancer cell lines MC38 (Cat# CL-0972) and CT26 (Cat# CL-0071), murine hepatocyte line AML12 (Cat# CL-0602), murine endothelial cell line SVEC4-10 (Cat# CL-0221), human umbilical vein endothelial cells (HUVEC, Cat# CL-0675), and human myeloid cell line HL-60 (Cat# CL-0110) were obtained from Procell Life Science & Technology Co., Ltd. (Wuhan, China).

Techniques: Injection, Control, Immunofluorescence, CCK-8 Assay, Functional Assay, Migration, Scratch Wound Assay Assay