Journal: Heliyon

Article Title: Exosomal CTHRC1 from cancer-associated fibroblasts facilitates endometrial cancer progression via ITGB3/FAK signaling pathway

doi: 10.1016/j.heliyon.2024.e35727

Figure Lengend Snippet: Abundant CTHRC1-loaded exosomes contribute EC cell migration via the activation of the ITGB3/FAK signaling pathway A Western blot analysis of CTHRC1 in HEK293 cells, transfected with lentiviral vectors to stable overexpress CTHRC1, empty lentivirus was used as negative control. B The presence of exosomal CTHRC1 and exosomal markers HSP70, CD63, and CD81 was detected by Western blotting. C,D The morphology and vesicle size distribution of exo-NC and exo-CTHRC1 were observed by TEM and NTA (TEM scale bar: 100 nm). E Exosomes (exo-NC, exo-CTHRC1) labeled with 1,10-dioctadecyl-3,3,30,30-tetramethylindocarbocyanine perchlorate (Dil, red) were taken up by Ishikawa and AN3CA cells (phalloidin-FITC in green, DAPI in blue), with PBS as negative control. F Cell proliferation following treatment with exo-CTHRC1 (25, 50 and 100 μg/ml) for 72 h as compared to exo-NC treatment were measured by cell counting kit-8 assays. G Cell migration of Ishikawa and AN3CA cells in response to exosomes (exo-NC, exo-CTHRC1,100 μg/ml) was assessed; scale bar: 100 μm (* p < 0.05). H The protein levels of ITGB3, p -FAK (Tyr397) and FAK after exo-CTHRC1 treatment (100 μg/ml) in EC cells were evaluated by Western blotting (GAPHD was used for normalization). I CTHRC1 was immunoprecipitated with ITGB3 antibody in Ishikawa and AN3CA cells after incubation with CTHRC1-enriched exosomes for 16 h. J, K EC cells were treated with exo-NC, exo-CTHRC1 and exo-CTHRC1+ Defactinib (inhibitor of FAK Tyr397 phosphorylation) for 16 h. Defactinib inhibited exosomal CTHRC1-induced phosphorylation of FAK (Tyr397) and cell migration in Ishikawa and AN3CA cells (* P < 0.05). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The membranes were blocked with 5 % bovine serum albumin for 2 h and incubated with antibodies against exosome-specific markers (CD9, CD63, and CD81; 1:1000, EXOAB-KIT-1, SBI, CA, USA), FAP (1:1000, ab53066, Abcam), α-SMA (1:1000, #19245, Cell Signalling Technology (CST), Chicago, USA), GAPDH (1:1000, #8884, CST), CTHRC1 (1 mg/ml, ab85739, Abcam), ITGB3 (1:1000, #13166, CST), p -FAK (1:1000, #8556, CST), and FAK (1:1000, #3285, CST) overnight at 4 °C, followed by incubation with HRP-conjugated secondary antibodies (1:2000, #7074,CST) for 2 h at room temperature.

Techniques: Migration, Activation Assay, Western Blot, Transfection, Negative Control, Labeling, Cell Counting, Immunoprecipitation, Incubation, Phospho-proteomics

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Enhancing endometrial receptivity: the roles of human chorionic gonadotropin in autophagy and apoptosis regulation in endometrial stromal cells

doi: 10.1186/s12958-024-01205-x

Figure Lengend Snippet: Endometrial receptivity is down-regulated in RIF patients. A The mRNA expression levels of HOXA10, ITGB3 and LHCGR in the endometrium of normal controls ( n = 12) and RIF patients ( n = 10). B WB was used to detect the protein levels of HOXA10, ITGB3 and LHCGR in endometrium of normal control ( n = 12) and RIF patients ( n = 10). Analyzing gels and western blots with ImageJ. Error bars, mean ± SD. * P < 0.05, ** P < 0.01. C Representative IHC images of LHCGR localization in endometrial tissue from normal control ( n = 12) and RIF patients ( n = 10) in secretory phase. Scale, 100 μm and 50 μm, analyzing relative OD with ImageJ. Error bars, mean ± SD. ** P < 0.01

Article Snippet: These membranes were saturated with blocking buffer for 1 h. Following this, membranes were incubated with rabbit polyclonal antiLHCGR (1:1,000 dilution; Proteintech), rabbit polyclonal anti-HOXA10 (1:1,000 dilution; Proteintech), rabbit polyclonal anti-ITGB3 antibodies (1:2,000 dilution; Proteintech), rabbit polyclonal anti-LIF antibodies (1:1,000 dilution; Proteintech) and mouse monoclonal anti-MECA-79 antibodies (1:500 dilution; Santa Cruz) at 4 °C.

Techniques: Expressing, Western Blot

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Enhancing endometrial receptivity: the roles of human chorionic gonadotropin in autophagy and apoptosis regulation in endometrial stromal cells

doi: 10.1186/s12958-024-01205-x

Figure Lengend Snippet: The expression levels of HOXA10, ITGB3, FOXO1, LIF and L-selectin ligand (MECA-79) in uterine ESCs after hCG intervention. A The mRNA expression levels of HOXA10, ITGB3, FOXO1 and LIF in ESCs were stimulated with hCG for 72 h. B The protein expression levels of HOXA10, ITGB3, FOXO1, LIF and L-selectin ligand (MECA-79) in ESCs were stimulated by hCG for 72 h. The data were shown as mean ± SD ( n = 3): * P < 0.05, ** P < 0.01, *** P < 0.001. C IF Representative images of ITGB3, LIF and L-selectin ligand (MECA-79) in ESCs treated with 0.1IU/mL hCG for 72 h. Scale: 100 μm

Article Snippet: These membranes were saturated with blocking buffer for 1 h. Following this, membranes were incubated with rabbit polyclonal antiLHCGR (1:1,000 dilution; Proteintech), rabbit polyclonal anti-HOXA10 (1:1,000 dilution; Proteintech), rabbit polyclonal anti-ITGB3 antibodies (1:2,000 dilution; Proteintech), rabbit polyclonal anti-LIF antibodies (1:1,000 dilution; Proteintech) and mouse monoclonal anti-MECA-79 antibodies (1:500 dilution; Santa Cruz) at 4 °C.

Techniques: Expressing

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Enhancing endometrial receptivity: the roles of human chorionic gonadotropin in autophagy and apoptosis regulation in endometrial stromal cells

doi: 10.1186/s12958-024-01205-x

Figure Lengend Snippet: The expression levels of LHCGR, FOXO1, ITGB3 ,HOXA10, LIF and L-selectin ligand in ESCs after the knockdown of LHCGR. A The mRNA expression levels of LCGR, HOXA10, ITGB3, FOXO1 and LIF in ESCs after the knockdown of LHCGR. B The protein expression levels of LHCGR in ESCs were stimulated by hCG and knockdown of LHCGR. C The protein expression levels of FOXO1, ITGB3, HOXA10, LIF and L-selectin ligand in ESCs after the knockdown of LHCGR. The data were shown as mean ± SD ( n = 3): * P < 0.05, ** P < 0.01

Article Snippet: These membranes were saturated with blocking buffer for 1 h. Following this, membranes were incubated with rabbit polyclonal antiLHCGR (1:1,000 dilution; Proteintech), rabbit polyclonal anti-HOXA10 (1:1,000 dilution; Proteintech), rabbit polyclonal anti-ITGB3 antibodies (1:2,000 dilution; Proteintech), rabbit polyclonal anti-LIF antibodies (1:1,000 dilution; Proteintech) and mouse monoclonal anti-MECA-79 antibodies (1:500 dilution; Santa Cruz) at 4 °C.

Techniques: Expressing

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Enhancing endometrial receptivity: the roles of human chorionic gonadotropin in autophagy and apoptosis regulation in endometrial stromal cells

doi: 10.1186/s12958-024-01205-x

Figure Lengend Snippet:

Article Snippet: These membranes were saturated with blocking buffer for 1 h. Following this, membranes were incubated with rabbit polyclonal antiLHCGR (1:1,000 dilution; Proteintech), rabbit polyclonal anti-HOXA10 (1:1,000 dilution; Proteintech), rabbit polyclonal anti-ITGB3 antibodies (1:2,000 dilution; Proteintech), rabbit polyclonal anti-LIF antibodies (1:1,000 dilution; Proteintech) and mouse monoclonal anti-MECA-79 antibodies (1:500 dilution; Santa Cruz) at 4 °C.

Techniques:

Journal: eLife

Article Title: Depletion of SMN protein in mesenchymal progenitors impairs the development of bone and neuromuscular junction in spinal muscular atrophy

doi: 10.7554/eLife.92731

Figure Lengend Snippet:

Article Snippet: Antibody , anti-Itgb3 (Rabbit monoclonal) , Cell Signaling , Cat. #: 13166; RRID: AB_2798136 , IF (1:100).

Techniques: Recombinant, Bicinchoninic Acid Protein Assay, Software, Staining