mouse monoclonal anti ena  (Developmental Studies Hybridoma Bank)

Journal: International journal of oncology

Article Title: High C-X-C motif chemokine 5 expression is associated with malignant phenotypes of prostate cancer cells via autocrine and paracrine pathways.

doi: 10.3892/ijo.2018.4388

Figure Lengend Snippet: Figure 1. CXCL5 expression in prostate cancer cells. (A) Reverse transcription-polymerase chain reaction results and semi-quantitative analysis of CXCL5 mRNA expression levels in PC‑3, DU145, LNCaP, WPMY‑1, and RWPE‑1 cell lines; β-actin was used to normalize the data; *P<0.05 and **P<0.01 vs. RWPE‑1 cells. (B) ELISA analysis of the concentrations of secreted CXCL5 proteins 48 h post-treatment in untransfected parental, mock-transfected, and CXCL5‑OE- transfected PC‑3, DU145 and WPMY‑1 cells, as well as LNCaP and RWPE‑1 cells. **P<0.01 vs. respective parental or mock cells. CXCL5, C-X-C motif chemokine 5; CXCL5‑OE, lentiviral-CXCL5-overexpression vector-transfected cells; mock, lentiviral empty vector-transfected cells.

Article Snippet: The sections were blocked with 20% normal goat serum (cat. no. AR0009; Wuhan Boster Biological Technology, Ltd.) at 37 ̊C for 1 h, followed by incubation with a mouse monoclonal anti-CXCL5 antibody (1:50; cat. no. sc‐73931; Santa Cruz Biotechnology, Inc.) at 4 ̊C overnight.

Techniques: Expressing, Reverse Transcription, Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Transfection, Over Expression, Plasmid Preparation

Journal: International journal of oncology

Article Title: High C-X-C motif chemokine 5 expression is associated with malignant phenotypes of prostate cancer cells via autocrine and paracrine pathways.

doi: 10.3892/ijo.2018.4388

Figure Lengend Snippet: Figure 2. Expression levels of CXCL5 protein in prostate cancer, cystic dilatation prostate, normal prostate and BPH tissue specimens. (A-I) Immunohistochemistry was performed to assess CXCL5 protein expression in the TMA and freshly collected tissue specimens; representative images are presented. (A-E) CXCL5 expression in TMA prostate cancer tissues with (A and B) Gleason grade 6, (C) Gleason grade 9, and (D and E) Gleason grade 10. (F) CXCL5 expression in fresh cystic dilatation prostate tissue. (G and H) CXCL5 expression in TMA normal prostate tissue. (I) CXCL5 expression in TMA BPH tissue. BPH, benign prostate hyperplasia; CXCL5, C-X-C motif chemokine 5; TMA, tissue microarray.

Article Snippet: The sections were blocked with 20% normal goat serum (cat. no. AR0009; Wuhan Boster Biological Technology, Ltd.) at 37 ̊C for 1 h, followed by incubation with a mouse monoclonal anti-CXCL5 antibody (1:50; cat. no. sc‐73931; Santa Cruz Biotechnology, Inc.) at 4 ̊C overnight.

Techniques: Expressing, Immunohistochemistry, Microarray

Journal: International journal of oncology

Article Title: High C-X-C motif chemokine 5 expression is associated with malignant phenotypes of prostate cancer cells via autocrine and paracrine pathways.

doi: 10.3892/ijo.2018.4388

Figure Lengend Snippet: Figure 3. Effects of exogenous CXCL5 protein exposure on prostate cancer cell proliferation, colony formation, and migration in vitro. (A) PC‑3 and DU145 cells were treated with various concentrations of recombinant CXCL5 (0, 10, 20, 40 and 60 ng/ml) for 72 h and proliferation was assessed using the CCK-8 assay. *P<0.05 and **P<0.01 vs. untreated (0 ng/ml) control cells. (B) PC‑3 and DU145 cells were treated with 60 ng/ml or 20 ng/ml recombinant CXCL5, respectively, for 12 days and colony formation was examined. (C) Transwell migration assay for PC‑3 and DU145 cells treated with various concentrations of CXCL5 (0, 10, 20, 40, and 60 ng/ml) for 48 h. (D) Results from statistical analysis of Transwell tumor cell migration assay from (C). **P<0.01 vs. 0, 10 or 60 ng/ ml; ##P<0.01 vs. 20 ng/ml; ▲▲P<0.01 vs. 0 or 60 ng/ml; &&P<0.01 vs. 40 ng/ml. CCK-8, Cell Counting Kit-8; CXCL5, C-X-C motif chemokine 5.

Article Snippet: The sections were blocked with 20% normal goat serum (cat. no. AR0009; Wuhan Boster Biological Technology, Ltd.) at 37 ̊C for 1 h, followed by incubation with a mouse monoclonal anti-CXCL5 antibody (1:50; cat. no. sc‐73931; Santa Cruz Biotechnology, Inc.) at 4 ̊C overnight.

Techniques: Migration, In Vitro, Recombinant, CCK-8 Assay, Control, Transwell Migration Assay, Cell Migration Assay, Cell Counting

Journal: International journal of oncology

Article Title: High C-X-C motif chemokine 5 expression is associated with malignant phenotypes of prostate cancer cells via autocrine and paracrine pathways.

doi: 10.3892/ijo.2018.4388

Figure Lengend Snippet: Figure 4. Effects of CXCL5 overexpression on prostate cancer cell proliferation and migration in vitro. (A) Cell proliferation of untransfected parental, mock- transfected, and CXCL5‑OE-transfected PC‑3 or DU145 cells was assessed using the CCK-8 assay. (B) Representative images of Transwell migration assay for parental, mock, and CXCL5‑OE PC‑3 and DU145 cells. (C) Analysis of cell migratory ability of PC‑3 and DU145 cells from (B). *P<0.05 and **P<0.01 vs. parental or mock. CCK-8, Cell Counting Kit-8; CXCL5, C-X-C motif chemokine 5; CXCL5‑OE, lentiviral-CXCL5-overexpression vector-transfected cells; mock, lentiviral empty vector-transfected cells.

Article Snippet: The sections were blocked with 20% normal goat serum (cat. no. AR0009; Wuhan Boster Biological Technology, Ltd.) at 37 ̊C for 1 h, followed by incubation with a mouse monoclonal anti-CXCL5 antibody (1:50; cat. no. sc‐73931; Santa Cruz Biotechnology, Inc.) at 4 ̊C overnight.

Techniques: Over Expression, Migration, In Vitro, Transfection, CCK-8 Assay, Transwell Migration Assay, Cell Counting, Plasmid Preparation

Journal: International journal of oncology

Article Title: High C-X-C motif chemokine 5 expression is associated with malignant phenotypes of prostate cancer cells via autocrine and paracrine pathways.

doi: 10.3892/ijo.2018.4388

Figure Lengend Snippet: Figure 5. Effects of CXCL5-overexpressing WPMY cells on prostate cancer cell malignant potential are paracrine in fashion. (A) PC‑3 and DU145 cells were cultured for 72 h in CM (20, 50, 80 or 100%) obtained from untransfected parental, mock-transfected, and CXCL5‑OE-transfected WPMY cells and proliferation was analyzed by CCK-8 assay. (B) Representative images of Transwell migration assay for PC‑3 and DU145 cells incubated with 100% parental, mock or CXCL5‑OE WPMY‑1 CM. (C) Analysis of migratory ability in PC‑3 and DU145 cells from (B). (D) Representative images of Transwell migration assay for PC‑3 and DU145 cells that were co-cultured with parental, mock, or CXCL5‑OE WPMY cells for 48 h. (E) Analysis of cell migratory ability of PC‑3 and DU145 cells from (D). *P<0.05 and **P<0.01 vs. parental or mock. CCK-8, Cell Counting Kit-8; CM, conditioned medium; CXCL5, C-X-C motif chemokine 5; CXCL5‑OE, lentiviral-CXCL5-overexpression vector-transfected cells; mock, lentiviral empty vector-transfected cells.

Article Snippet: The sections were blocked with 20% normal goat serum (cat. no. AR0009; Wuhan Boster Biological Technology, Ltd.) at 37 ̊C for 1 h, followed by incubation with a mouse monoclonal anti-CXCL5 antibody (1:50; cat. no. sc‐73931; Santa Cruz Biotechnology, Inc.) at 4 ̊C overnight.

Techniques: Cell Culture, Transfection, CCK-8 Assay, Transwell Migration Assay, Incubation, Cell Counting, Over Expression, Plasmid Preparation

Journal: International journal of oncology

Article Title: High C-X-C motif chemokine 5 expression is associated with malignant phenotypes of prostate cancer cells via autocrine and paracrine pathways.

doi: 10.3892/ijo.2018.4388

Figure Lengend Snippet: Figure 6. Effects of CXCL5 gene expression in PC‑3 prostate cancer cells. Untransfected parental, mock-transfected, or CXCL5‑OE-transfected PC‑3 cells were used to determine the effects of CXCL5 on the (A and B) mRNA and (C and D) protein expression levels of BAX, NDRG3, ERK, and CXCR2. *P<0.05 and **P<0.01 vs. parental or mock. CXCL5, C-X-C motif chemokine 5; ERK, extracellular signal-regulated kinase; NDRG3, N-Myc downstream-regulated gene 3. CXCR2, C-X-C chemokine receptor type 2; CXCL5‑OE, lentiviral-CXCL5-overexpression vector-transfected cells; mock, lentiviral empty vector-transfected cells.

Article Snippet: The sections were blocked with 20% normal goat serum (cat. no. AR0009; Wuhan Boster Biological Technology, Ltd.) at 37 ̊C for 1 h, followed by incubation with a mouse monoclonal anti-CXCL5 antibody (1:50; cat. no. sc‐73931; Santa Cruz Biotechnology, Inc.) at 4 ̊C overnight.

Techniques: Gene Expression, Transfection, Expressing, Over Expression, Plasmid Preparation

Journal: International journal of oncology

Article Title: High C-X-C motif chemokine 5 expression is associated with malignant phenotypes of prostate cancer cells via autocrine and paracrine pathways.

doi: 10.3892/ijo.2018.4388

Figure Lengend Snippet: Figure 7. Effects of CXCL5 overexpression on PC‑3 prostate cancer cell‑derived xenograft formation and growth in nude mice. Mock-transfected or CXCL5‑OE- transfected PC‑3 cells (1x106) were subcutaneously injected into nude mice; the mice were sacrificed after 8 weeks and the tumors were analyzed for (A) sizes and (B) weight. (C) Tumor cell xenografts were resected and subjected to western blot analysis for IL‑18, Bcl‑2 and caspase‑3 protein expression levels. (D) Quantified data from the western blot analysis in (C). *P<0.05 and **P<0.01 vs. mock. CXCL5, C-X-C motif chemokine 5; CXCL5‑OE, lentiviral-CXCL5- overexpression vector-transfected cells; IL, interleukin; mock, lentiviral empty vector-transfected cells.

Article Snippet: The sections were blocked with 20% normal goat serum (cat. no. AR0009; Wuhan Boster Biological Technology, Ltd.) at 37 ̊C for 1 h, followed by incubation with a mouse monoclonal anti-CXCL5 antibody (1:50; cat. no. sc‐73931; Santa Cruz Biotechnology, Inc.) at 4 ̊C overnight.

Techniques: Over Expression, Transfection, Injection, Western Blot, Expressing, Plasmid Preparation