Journal: Clinical & experimental metastasis

Article Title: EGFL6 promotes bone metastasis of lung adenocarcinoma by increasing cancer cell malignancy and bone resorption.

doi: 10.1007/s10585-023-10219-5

Figure Lengend Snippet: Fig. 1 EGFL6 is upregulated in lung adenocarcinoma tissues. The images of EGFL6 IHC staining in lung adenocarcinoma tissues and adjacent matched tissues. Scale bar represents 100 μm

Article Snippet: EGFL6 protein (MCE, China), derived from mouse, was added into the rEGFL6 group at the concentration of 1.33 mg/μl according to the manufacturer’s instructions.

Techniques: Immunohistochemistry

Journal: Clinical & experimental metastasis

Article Title: EGFL6 promotes bone metastasis of lung adenocarcinoma by increasing cancer cell malignancy and bone resorption.

doi: 10.1007/s10585-023-10219-5

Figure Lengend Snippet: Fig. 3 Effects of EGFL6 on EMT- related gene expression. (a) RT- PCR shows that overexpression of EGFL6 promotes the expres sion of N-cadherin, MMP2, MMP9, Vimentin, Snail, and Slug and down-regulates the expres sion of E-cadherin. Knockdown of EGFL6 exhibits the opposite results. (b,c) Western blot repli cates the RT-PCR results. (d,e) The effect of EGFL6 on EMT markers (E-cadherin, N-cadherin, and Vimentin) in A549 cells is detected by ICC. Over-expression of EGFL6 increases the expres sion of N-cadherin and Vimentin in cytoplasm but attenuates the expression of E-cadherin while knockdown of EGFL6 dis plays the adverse. Results were presented as mean ± SD. P < 0.05 was considered statistically significant. *P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: EGFL6 protein (MCE, China), derived from mouse, was added into the rEGFL6 group at the concentration of 1.33 mg/μl according to the manufacturer’s instructions.

Techniques: Gene Expression, Reverse Transcription Polymerase Chain Reaction, Over Expression, Knockdown, Western Blot, Expressing

Journal: Clinical & experimental metastasis

Article Title: EGFL6 promotes bone metastasis of lung adenocarcinoma by increasing cancer cell malignancy and bone resorption.

doi: 10.1007/s10585-023-10219-5

Figure Lengend Snippet: Fig. 5 EGFL6 moderates the effect of A549 cells on skel eton system of nude mice and enhances the osteoclast differ entiation of BMMs. (a,b) The tumor size in the H-EGFL6 group is much larger than the H-Ctrl group, and the L-EGFL6 group exhibited a smaller size than the L-Ctrl group. (c) Compared with each control, X-ray shows that the bone destruction of tibia in H-EGFL6 group is much severe, while in L-EGFL6 group is slight. (d) Histomorphometric quantitative analysis of per centage bone volume to tissue volume (BV/TV, %). (e,f) The tibial tumor sections of nude mice were stained with H&E. (g) The tibial tumor sections of nude mice were stained with TRAP. (h) The number of TRAP- positive osteoclasts per bone surface (OCs/BS). Results were presented as mean ± SD. P < 0.05 was considered statistically significant. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: EGFL6 protein (MCE, China), derived from mouse, was added into the rEGFL6 group at the concentration of 1.33 mg/μl according to the manufacturer’s instructions.

Techniques: Control, Staining

Journal: Clinical & experimental metastasis

Article Title: EGFL6 promotes bone metastasis of lung adenocarcinoma by increasing cancer cell malignancy and bone resorption.

doi: 10.1007/s10585-023-10219-5

Figure Lengend Snippet: Fig. 6 EGFL6 promotes osteo clast differentiation of BMMs but has no significant effect on osteo blast differentiation of BMSCs as an exocrine protein. (a) ELISA assay shows that EGFL6 is an exocrine protein. (b,c) EGFL6 has no significant influence on osteoblast differentiation of BMSCs. (d,e) EGFL6 enhances the osteoclast differentiation of BMMs. (f,g) EGFL6 protein promotes the osteoclast differen tiation. (h,i) The bone resorption pits assay. Results were presented as mean ± SD. P < 0.05 was con sidered statistically significant. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: EGFL6 protein (MCE, China), derived from mouse, was added into the rEGFL6 group at the concentration of 1.33 mg/μl according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Molecular Medicine Reports

Article Title: Characterization of bone marrow-derived mesenchymal stem cells from dimethyloxallyl glycine-preconditioned mice: Evaluation of the feasibility of dimethyloxallyl glycine as a mobilization agent

doi: 10.3892/mmr.2016.4945

Figure Lengend Snippet: Paracrine capacity of NBM-MSCs and DBM-MSCs. (A) Quantification of TGF concentration in the culture supernatant of NBM-MSCs and DBM-MSCs. There was no significant difference between the expression levels (P>0.05). (B) Quantification of PDGF concentration in the culture supernatant of NBM-MSCs and DBM-MSCs. No significant difference was identified between them (P>0.05). TGF and PDGF concentrations were detected using enzyme-linked immunosorbent assays. Data are presented as the mean ± standard deviation. BM-MSCs, bone marrow-derived mesenchymal stem cells from normal saline-treated mice; DBM-MSCs, bone marrow-derived mesenchymal stem cells from dimethyloxallyl glycine-preconditioned mice; TGF, transforming growth factor; PDGF, platelet-derived growth factor.

Article Snippet: Mouse TGF (β IG-H3) and PDGF ELISA kits (Wuhan Boster Biological Co., Ltd., Wuhan, China) were used, according to the manufacturer's protocol.

Techniques: Concentration Assay, Expressing, Standard Deviation, Derivative Assay, Saline

Journal: Discover Oncology

Article Title: Crosstalk between cancer cells and macrophages promotes OSCC cell migration and invasion through a CXCL1/EGF positive feedback loop

doi: 10.1007/s12672-024-00972-8

Figure Lengend Snippet: CXCL1 induces OSCC cell migration and invasion via macrophage crosstalk. A Measurement of CXCL1 mRNA expression by RT‒PCR in Hacat cells and Cal27 cells. B Secretion levels of CXCL1 in Hacat cells and Cal27 cells were measured by ELISA. C Wound healing assays showed the migration of Cal27 cells. D Transwell assays showed the migratory and invasive abilities of Cal27 cells. Bar graphs are presented as the mean ± SD (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 vs. the control

Article Snippet: Measurement of EGF and CXCL1 levels in the supernatant of the TAMs and Cal27 cells was conducted using mouse EGF ELISA kits (BOSTER Biological Technology Co. Ltd., Wuhan, China; EK0326) and human CXCL1 ELISA kits (R&D Systems Inc., Minneapolis, MN, USA; cat. no. #DY275), respectively.

Techniques: Migration, Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Discover Oncology

Article Title: Crosstalk between cancer cells and macrophages promotes OSCC cell migration and invasion through a CXCL1/EGF positive feedback loop

doi: 10.1007/s12672-024-00972-8

Figure Lengend Snippet: M2 macrophage polarization is enhanced and EGF expression is upregulated in macrophages. A – C Gene expression of CD206, CD86, and Arg-1 was measured by RT‒PCR. D – G Flow cytometric analysis of surface markers of CD206 and CD86 in RAW264.7 cells incubated with CXCL1 (100 ng/ml) or without CXCL1-containing tumor-conditioned medium. H , I mRNA and protein expression levels of EGF in RAW264.7 cells incubated with or without CXCL1 (100 ng/ml) in tumor-conditioned medium. J , K mRNA and protein expression of EGF in RAW264.7 cells incubated with Cal27-transfected supernatant or Cal27-CM. L RT‒PCR was used to examine the mRNA expression of EGF in RAW264.7 cells. M Immunosorbent assay (ELISA) showing the protein level of EGF in RAW264.7 cells. Bar graphs are presented as the mean ± SD (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 vs. the control

Article Snippet: Measurement of EGF and CXCL1 levels in the supernatant of the TAMs and Cal27 cells was conducted using mouse EGF ELISA kits (BOSTER Biological Technology Co. Ltd., Wuhan, China; EK0326) and human CXCL1 ELISA kits (R&D Systems Inc., Minneapolis, MN, USA; cat. no. #DY275), respectively.

Techniques: Expressing, Gene Expression, Incubation, Transfection, Enzyme-linked Immunosorbent Assay, Control

Journal: Discover Oncology

Article Title: Crosstalk between cancer cells and macrophages promotes OSCC cell migration and invasion through a CXCL1/EGF positive feedback loop

doi: 10.1007/s12672-024-00972-8

Figure Lengend Snippet: EGF promotes OSCC progression by activating the EGFR/NF-κB pathway. A Cal27 cells were treated with EGF (100 ng/ml) for 0, 30, 60, and 120 min, and the expression of EGFR was detected by Western blot analysis. B , C Quantified detection of the EGFR expression. D Western blot analysis of the expression of EGFR and members of the NF-κB signaling pathway in Cal27 cells treated with EGF or EGF and AG1478. E , F Quantitative analysis of Western blots in ( D ) Cal27 cells. G mRNA expression of CXCL1 in Cal27 cells treated with EGF (100 ng/ml) or EGF and AG1478 (5 μM) in TAM-CM. H Measurement of CXCL1 levels in treated Cal27 cells by ELISA. I Western blot analysis of the expression of CXCL1 in Cal27 cells treated with EGF or EGF and AG1478-containing TAM-CM. Bar graphs are presented as the mean ± SD (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001 vs. the control

Article Snippet: Measurement of EGF and CXCL1 levels in the supernatant of the TAMs and Cal27 cells was conducted using mouse EGF ELISA kits (BOSTER Biological Technology Co. Ltd., Wuhan, China; EK0326) and human CXCL1 ELISA kits (R&D Systems Inc., Minneapolis, MN, USA; cat. no. #DY275), respectively.

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Control