Journal: Journal of Cellular and Molecular Medicine

Article Title: Occludin facilitates tumour angiogenesis in bladder cancer by regulating IL8/STAT3 through STAT4

doi: 10.1111/jcmm.17257

Figure Lengend Snippet: Occludin (OCLN) promotes tumour angiogenesis in vitro and in vivo. (a and b) The knockdown efficiency was confirmed by performing (A) Western blotting and (B) RT‐qPCR assays using 5637 and T24 cells. (C), Tube formation by EA.hy926 cells incubated with CM derived from 5637 and T24 OCLN‐silenced bladder cancer (BLCA) cells was assessed using staining with Calcein AM and imaging with a fluorescence microscope. Scale bar = 100 µm. (d and e) The OCLN plasmid was transfected into 5637 and T24 cells; the efficiency of overexpression was analysed using (D) Western blotting and (E) RT‐qPCR. (F) Tube formation by EA.hy926 cells incubated with CM derived from 5637 and T24 OCLN overexpressing cells was assessed using staining with Calcein AM and imaging with a fluorescence microscope. Scale bar = 100 µm. (G) The segment lengths were analysed, and the meshes were quantified using ImageJ software. (H) Matrigel plugs containing 5637 and T24 OCLN stable knockdown cells were removed from BALB/c nude mice. (I) CD31 staining in the indicated cells embedded in Matrigel plugs after growth in BALB/c nude mice. Scale bar = 100 µm. (J) The density of microvessels in Matrigel plugs from BALB/c nude mice injected with the indicated cells. (K) IHC staining showing CD31 levels in clinical patients with high‐/low‐grade BLCA. Scale bar = 100 µm. The results are shown as the mean ± SD. *.01 < p < .05; **.001 < p < .01

Article Snippet: BLCA cell lines, namely T24 (ATCC HTB‐4), 5637 (ATCC HTB‐9) were derived from laboratory retained cells.

Techniques: In Vitro, In Vivo, Knockdown, Western Blot, Quantitative RT-PCR, Incubation, Derivative Assay, Staining, Imaging, Fluorescence, Microscopy, Plasmid Preparation, Transfection, Over Expression, Software, Injection, Immunohistochemistry

Journal: Journal of Cellular and Molecular Medicine

Article Title: Occludin facilitates tumour angiogenesis in bladder cancer by regulating IL8/STAT3 through STAT4

doi: 10.1111/jcmm.17257

Figure Lengend Snippet: Occludin (OCLN) mediates bladder cancer (BLCA) angiogenesis by regulating IL8 expression. (A) Venn diagram showing the differentially expressed genes (DEGs) in the two T24 OCLN knockdown groups compared with the control groups (fold change ≥1, FDR < 0.1, p < .05). (B) Heatmap of the RNA sequencing analysis showing the relative levels of proangiogenic factors. Columns represent probe sets, and rows represent samples receiving the indicated treatments. (C) The relative mRNA levels of proangiogenic factors were detected in control and OCLN shRNA transfected 5637 and T24 cells. (D) The relative IL8 levels in control and OCLN shRNA‐transfected 5637 and T24 cells were measured using an ELISA (pg/ml). (E) The relative IL8 mRNA levels were detected in 5637 and T24 cells following transfection with the vector or OCLN plasmid. (F) Tube formation by EA.hy926 cells cultured with CM derived from 5637 and T24 OCLN‐silenced BLCA cells. IL8 was added, and the cells were stained with Calcein AM and then imaged with a fluorescence microscope. Scale bar = 100 µm. (G) The segment lengths were analysed, and the meshes were quantified using ImageJ software. The results are shown as the mean ± SD. *.01 < p < .05; **.001 < p < .01; *** p < .001

Article Snippet: BLCA cell lines, namely T24 (ATCC HTB‐4), 5637 (ATCC HTB‐9) were derived from laboratory retained cells.

Techniques: Expressing, Knockdown, Control, RNA Sequencing, shRNA, Transfection, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Cell Culture, Derivative Assay, Staining, Fluorescence, Microscopy, Software

Journal: Journal of Cellular and Molecular Medicine

Article Title: Occludin facilitates tumour angiogenesis in bladder cancer by regulating IL8/STAT3 through STAT4

doi: 10.1111/jcmm.17257

Figure Lengend Snippet: IL8/ STAT3 is involved in the process of Occludin (OCLN)‐mediated angiogenesis in bladder cancer (BLCA). (A) p‐STAT3 and STAT3 protein levels were detected in OCLN knockdown 5637 and T24 cells. (B) 5637 and T24 cells were transfected with the OCLN plasmid and treated with or without the STAT3 inhibitor Stattic, and tube formation by EA.hy926 cells incubated with CM derived from the indicated cells was assessed by performing staining with Calcein AM and imagining using a fluorescence microscope. Scale bar = 100 µm. (C) The segment lengths in these images were analysed, and the meshes were quantified using ImageJ software. D, p‐STAT3 and STAT3 protein levels were detected in OCLN knockdown 5637 and T24 cells after IL8 supplementation. (E) p‐STAT3 and STAT3 protein levels were detected in OCLN‐overexpressing 5637 and T24 cells after blocking IL8. (F) 5637 and T24 cells were transfected with the OCLN plasmid or cultured with the IL8‐neutralizing antibody; tube formation by EA.hy926 cells incubated with CM derived from the indicated cells was assessed using staining with Calcein AM and imaging with a fluorescence microscope. Scale bar = 100 µm. G, The segment lengths in these images were analysed, and the meshes were quantified using ImageJ software. The results are shown as the mean ± SD. *.01< p < .05; **.001< p < .01

Article Snippet: BLCA cell lines, namely T24 (ATCC HTB‐4), 5637 (ATCC HTB‐9) were derived from laboratory retained cells.

Techniques: Knockdown, Transfection, Plasmid Preparation, Incubation, Derivative Assay, Staining, Fluorescence, Microscopy, Software, Blocking Assay, Cell Culture, Imaging

Journal: Cureus

Article Title: Elevated Plasma Levels of CXCL13 Chemokine in Saudi Patients With Asthma Exacerbation

doi: 10.7759/cureus.21142

Figure Lengend Snippet: CXCL13 protein concentration was determined in the plasma by ELISA assay. Data was presented as mean ± SD (normal = 20, asthma = 23, P < 0.0001). ELISA, Enzyme-linked immunoassay.

Article Snippet: Quantification of total IgE and CXCL13 protein by ELISA Total IgE (Cat: ab108650, Abcam plc, Cambridge, UK) and CXCL13 chemokine (Cat: DY801, R&D, UK) levels were quantified in the plasma of the asthma group and healthy controls as described by the manufacturer.

Techniques: Protein Concentration, Enzyme-linked Immunosorbent Assay