rabbit anti human cxcr7 antibody  (Proteintech)

Journal: Oncology reports

Article Title: Overexpression of CXCR7 induces angiogenic capacity of human hepatocellular carcinoma cells via the AKT signaling pathway.

doi: 10.3892/or.2016.5045

Figure Lengend Snippet: Figure 1. The CXCR7 expression in different metastatic HCC cell lines. (A) The mRNA expression of CXCR7 was examined by real‑time PCR. (B) The protein expression of CXCR7 was determined by western blotting. Data are means ± SD of three independent experiments. *P<0.05. CXCR7, C‑X‑C chemokine receptor type 7; HCC, hepatocellular carcinoma.

Article Snippet: Rabbit anti-human CXCR7 antibody (1:200; R&D Systems, Inc., Minneapolis, MN, USA) followed by 1:3,000 horseradish peroxidase-conjugated goat anti-rabbit IgG F(ab')2 antibody (Jackson ImmunoResearch, West Grove, PA, USA) was used.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Oncology reports

Article Title: Overexpression of CXCR7 induces angiogenic capacity of human hepatocellular carcinoma cells via the AKT signaling pathway.

doi: 10.3892/or.2016.5045

Figure Lengend Snippet: Figure 2. Overexpression of CXCR7 induces angiogenic capacity of HCC cells in vitro. (A) Western blotting of HCCLM3‑vector, HCCLM3‑CXCR7 protein expression in HCCLM3 cells; α‑tubulin was used as a control. The numbers show the relative level of protein compared to the control. (B) The images (left) and quantification (right) of tube formation by HUVECs cultured in CM. (C) The migration images and quantification of HUVECs after incubation in CM. (D) The images (left) and quantification (right) of blood vessels in the CAM assay when stimulated by CM. Data are means ± SD of three independent experiments. *P<0.05. CXCR7, C‑X‑C chemokine receptor type 7; HCC, hepatocellular carcinoma; HUVECs, human umbilical vein endothelial cells; CM, conditioned medium; CAM, chicken chorioallantoic membrane.

Article Snippet: Rabbit anti-human CXCR7 antibody (1:200; R&D Systems, Inc., Minneapolis, MN, USA) followed by 1:3,000 horseradish peroxidase-conjugated goat anti-rabbit IgG F(ab')2 antibody (Jackson ImmunoResearch, West Grove, PA, USA) was used.

Techniques: Over Expression, In Vitro, Western Blot, Expressing, Control, Cell Culture, Migration, Incubation, Chick Chorioallantoic Membrane Assay, Membrane

Journal: Oncology reports

Article Title: Overexpression of CXCR7 induces angiogenic capacity of human hepatocellular carcinoma cells via the AKT signaling pathway.

doi: 10.3892/or.2016.5045

Figure Lengend Snippet: Figure 3. Knockdown of CXCR7 decreases the angiogenic capacity of HCC cells in vitro. (A) Western blotting of CXCR7‑shRNA‑transduced HCCLM3 protein expression in HCCLM3 cells; α‑tubulin was used as a control. The numbers show the relative level of protein compared to the control. (B) The images (left) and quantification (right) of tube formation by HUVECs cultured in CM. (C) The migration images and quantification of HUVECs after incubation in CM. (D) The images (left) and quantification (right) of blood vessels in the CAM assay when stimulated by CM. Data are means ± SD of three independent experiments. *P<0.05. CXCR7, C‑X‑C chemokine receptor type 7; HCC, hepatocellular carcinoma; HUVECs, human umbilical vein endothelial cells; CM, conditioned medium; CAM, chicken chorioallantoic membrane.

Article Snippet: Rabbit anti-human CXCR7 antibody (1:200; R&D Systems, Inc., Minneapolis, MN, USA) followed by 1:3,000 horseradish peroxidase-conjugated goat anti-rabbit IgG F(ab')2 antibody (Jackson ImmunoResearch, West Grove, PA, USA) was used.

Techniques: Knockdown, In Vitro, Western Blot, Expressing, Control, Cell Culture, Migration, Incubation, Chick Chorioallantoic Membrane Assay, Membrane

Journal: Oncology reports

Article Title: Overexpression of CXCR7 induces angiogenic capacity of human hepatocellular carcinoma cells via the AKT signaling pathway.

doi: 10.3892/or.2016.5045

Figure Lengend Snippet: Figure 5. Overexpression of CXCR7 enhances the angiogenic capacity of HCC cells via activating the AKT pathway. CXCR7‑overexpressing HCC cells were inhibited with LY294002, which uses as a specific AKT inhibitor. (A) The images and quantification of tube formation by HUVECs on CM from the indicated cells. (B) The migration images and quantification of HUVECs after incubation in CM. (C) The images and quantification of blood vessels in the CAM assay when stimulated by CM. (D) ELISA of TNF‑α, IL‑6 and IL‑8 protein expression in the indicated cells *P<0.05. CXCR7, C‑X‑C chemokine receptor type 7; HCC, hepatocellular carcinoma; HUVECs, human umbilical vein endothelial cells; CM, conditioned medium; CAM, chicken chorioallantoic membrane; ELISA, enzyme-linked immunosorbent assay; TNF, tumor necrosis factor; IL, interleukin.

Article Snippet: Rabbit anti-human CXCR7 antibody (1:200; R&D Systems, Inc., Minneapolis, MN, USA) followed by 1:3,000 horseradish peroxidase-conjugated goat anti-rabbit IgG F(ab')2 antibody (Jackson ImmunoResearch, West Grove, PA, USA) was used.

Techniques: Over Expression, Migration, Incubation, Chick Chorioallantoic Membrane Assay, Enzyme-linked Immunosorbent Assay, Expressing, Membrane

Journal: Oncology reports

Article Title: CXCR7 correlates with the differentiation of hepatocellular carcinoma and suppresses HNF4α expression through the ERK pathway.

doi: 10.3892/or.2014.3501

Figure Lengend Snippet: Figure 3. CXCR7 is inversely correlated to HNF4α upon induced differentia- tion. (A) Huh7 and HCCLM3 cells were treated with 1% DMSO, respectively. The expression level of CXCR7 was elevated, and the HNF4α level was decreased during the early stage of induced differentiation as observed by western blotting. During the late stage of induced differentiation, the level of CXCR7 was decreased and the HNF4α level was elevated. (B) HCCLM3 cells were treated with OSM. The CXCR7 level was elevated while the oppo- site trend was observed with HNF4α during the early stage of differentiation in response to 40 ng/ml OSM treatment.

Article Snippet: Antibodies used for immunofluorescence, immunoblotting and immunohistochemistry were as follows: mouse anti-human monoclonal CD90 (Abcam, Cambridge, MA, USA), rabbit anti-human polyclonal CD133 (Abnova, Walnut, CA, USA), mouse anti-human monoclonal CD44, rabbit anti-human albumin (ALB), phospho-p44/42 MAPK (Erk1/2), and rabbit anti-human HnF4α mAb (Cell Signaling Technologies, Danvers, MA, USA), rabbit anti-human polyclonal CD24, rabbit anti-human β-actin mAb (Epitomics, Burlingame, CA, USA), rabbit anti-human polyclonal transferrin (TF) (Proteintech Group Inc., Chicago, IL, USA), rabbit anti-human AFP mAb, rabbit anti-human C/EBPα and C/ EBPβ mAb, Erk2/p42 MAPK antibody (Epitomics), rabbit anti-human CXCR7 Igg (novus Biologicals, Littleton, CO, USA), and horseradish peroxidase-conjugated goat anti-rabbit IgG F(ab')2 antibodies (Jackson ImmunoResearch, West Grove, PA, USA).

Techniques: Expressing, Western Blot

Journal: Oncology reports

Article Title: CXCR7 correlates with the differentiation of hepatocellular carcinoma and suppresses HNF4α expression through the ERK pathway.

doi: 10.3892/or.2014.3501

Figure Lengend Snippet: Figure 4. Immunostaining of CXCR7 and HNF4α in the TMA (A) Weak or strong staining was recognized as low or high CXCR7 expression, which mainly stained in the cytoplasm or on the surface of the tumor cells. (B) Low and high HNF4α levels are shown. The staining of HNF4α was mainly nuclear. The posi- tive staining in normal hepatocytes indicated the specificity of the HNF4α antibody. (C) IHC analysis in representative cases indicated that tumor tissues with high CXCR7 expression always had low HNF4α expression, whereas CXCR7Low tissues were also likely to be HNF4αHigh, particularly in the well-differentiated HCC cases. (x200 original magnification; scale bar, 100 µm).

Article Snippet: Antibodies used for immunofluorescence, immunoblotting and immunohistochemistry were as follows: mouse anti-human monoclonal CD90 (Abcam, Cambridge, MA, USA), rabbit anti-human polyclonal CD133 (Abnova, Walnut, CA, USA), mouse anti-human monoclonal CD44, rabbit anti-human albumin (ALB), phospho-p44/42 MAPK (Erk1/2), and rabbit anti-human HnF4α mAb (Cell Signaling Technologies, Danvers, MA, USA), rabbit anti-human polyclonal CD24, rabbit anti-human β-actin mAb (Epitomics, Burlingame, CA, USA), rabbit anti-human polyclonal transferrin (TF) (Proteintech Group Inc., Chicago, IL, USA), rabbit anti-human AFP mAb, rabbit anti-human C/EBPα and C/ EBPβ mAb, Erk2/p42 MAPK antibody (Epitomics), rabbit anti-human CXCR7 Igg (novus Biologicals, Littleton, CO, USA), and horseradish peroxidase-conjugated goat anti-rabbit IgG F(ab')2 antibodies (Jackson ImmunoResearch, West Grove, PA, USA).

Techniques: Immunostaining, Staining, Expressing

Journal: Oncology reports

Article Title: CXCR7 correlates with the differentiation of hepatocellular carcinoma and suppresses HNF4α expression through the ERK pathway.

doi: 10.3892/or.2014.3501

Figure Lengend Snippet: Figure 5. CXCR7 is a strong prognostic biomarker for poor survival when combined with low HNF4α expression. In HNF4αLow HCC, high CXCR7 expres- sion was correlated with (A) poor overall survival and (B) reduced time to progression, particularly during the early stages of follow-up. (C) In addition, extrahepatic metastases occurred more rapidly in patients with CXCR7High HCC. Further stratification based on relative degree of differentiation indicated the prognostic value of CXCR7 in the survival of patients post-resection, including (D) overall survival, (E) time to progression and (F) time to metastasis.

Article Snippet: Antibodies used for immunofluorescence, immunoblotting and immunohistochemistry were as follows: mouse anti-human monoclonal CD90 (Abcam, Cambridge, MA, USA), rabbit anti-human polyclonal CD133 (Abnova, Walnut, CA, USA), mouse anti-human monoclonal CD44, rabbit anti-human albumin (ALB), phospho-p44/42 MAPK (Erk1/2), and rabbit anti-human HnF4α mAb (Cell Signaling Technologies, Danvers, MA, USA), rabbit anti-human polyclonal CD24, rabbit anti-human β-actin mAb (Epitomics, Burlingame, CA, USA), rabbit anti-human polyclonal transferrin (TF) (Proteintech Group Inc., Chicago, IL, USA), rabbit anti-human AFP mAb, rabbit anti-human C/EBPα and C/ EBPβ mAb, Erk2/p42 MAPK antibody (Epitomics), rabbit anti-human CXCR7 Igg (novus Biologicals, Littleton, CO, USA), and horseradish peroxidase-conjugated goat anti-rabbit IgG F(ab')2 antibodies (Jackson ImmunoResearch, West Grove, PA, USA).

Techniques: Biomarker Discovery, Expressing

Journal: Oncology reports

Article Title: CXCR7 correlates with the differentiation of hepatocellular carcinoma and suppresses HNF4α expression through the ERK pathway.

doi: 10.3892/or.2014.3501

Figure Lengend Snippet: Figure 6. HCCLM3 is a CXCR7+CXCR3-CXCR4- HCC cell line. Human chemokine and chemokine receptor GEArray kits were hybridized with cDNA probes derived from HCCLM3, MHCC97-L and SMMC-7721 cells. Strong CXCR7 and non-detectable CXCR3 and CXCR4 signals were observed for the HCCLM3 cell line.

Article Snippet: Antibodies used for immunofluorescence, immunoblotting and immunohistochemistry were as follows: mouse anti-human monoclonal CD90 (Abcam, Cambridge, MA, USA), rabbit anti-human polyclonal CD133 (Abnova, Walnut, CA, USA), mouse anti-human monoclonal CD44, rabbit anti-human albumin (ALB), phospho-p44/42 MAPK (Erk1/2), and rabbit anti-human HnF4α mAb (Cell Signaling Technologies, Danvers, MA, USA), rabbit anti-human polyclonal CD24, rabbit anti-human β-actin mAb (Epitomics, Burlingame, CA, USA), rabbit anti-human polyclonal transferrin (TF) (Proteintech Group Inc., Chicago, IL, USA), rabbit anti-human AFP mAb, rabbit anti-human C/EBPα and C/ EBPβ mAb, Erk2/p42 MAPK antibody (Epitomics), rabbit anti-human CXCR7 Igg (novus Biologicals, Littleton, CO, USA), and horseradish peroxidase-conjugated goat anti-rabbit IgG F(ab')2 antibodies (Jackson ImmunoResearch, West Grove, PA, USA).

Techniques: Derivative Assay

Journal: Oncology reports

Article Title: CXCR7 correlates with the differentiation of hepatocellular carcinoma and suppresses HNF4α expression through the ERK pathway.

doi: 10.3892/or.2014.3501

Figure Lengend Snippet: Figure 7. CXCR7 signaling suppresses HNF4α expression in an ERK-dependent fashion in HCC. (A) HCCLM3 and MHCC97H cells were cultured with or without the MEK1/2 inhibitor U0126. Phosphorylation of MAPKs ERK1/2 was decreased at 1, 8 or 16 h, and was accompanied by increased HNF4α expression, which indicates the suppression of the ERK pathway by HNF4α. (B) Cells were cultured in the presence of CXCR7 ligands (100 ng/ml SDF-1α or 200 ng/ml CXCL11). Elevated phosphorylation of ERK1/2 was observed a short time after ligand activation, which indicated that CXCR7 ligands can activate ERK signaling. (C) HCCLM3 and MHCC97H cells were cultured in the presence of CXCR7 ligands (100 ng/ml SDF-1α or 200 ng/ml CXCL11) with or without the MEK1/2 inhibitor U0126. Western blotting indicated the elevated HNF4α level after using U0126, indicating that the use of U0126 abrogated the inhibitory effect of CXCR7 ligands on HNF4α expression. (D) Knockdown of CXCR7 increased HNF4α expression in HCC cells. C/EBPα, another critical transcriptional factor, did not exhibit substantial changes. (E) Schematic drawing illustrating the proposed mechanism by which the transmembrane receptor CXCR7 mediates the ERK-dependent pathway to suppress HNF4α

Article Snippet: Antibodies used for immunofluorescence, immunoblotting and immunohistochemistry were as follows: mouse anti-human monoclonal CD90 (Abcam, Cambridge, MA, USA), rabbit anti-human polyclonal CD133 (Abnova, Walnut, CA, USA), mouse anti-human monoclonal CD44, rabbit anti-human albumin (ALB), phospho-p44/42 MAPK (Erk1/2), and rabbit anti-human HnF4α mAb (Cell Signaling Technologies, Danvers, MA, USA), rabbit anti-human polyclonal CD24, rabbit anti-human β-actin mAb (Epitomics, Burlingame, CA, USA), rabbit anti-human polyclonal transferrin (TF) (Proteintech Group Inc., Chicago, IL, USA), rabbit anti-human AFP mAb, rabbit anti-human C/EBPα and C/ EBPβ mAb, Erk2/p42 MAPK antibody (Epitomics), rabbit anti-human CXCR7 Igg (novus Biologicals, Littleton, CO, USA), and horseradish peroxidase-conjugated goat anti-rabbit IgG F(ab')2 antibodies (Jackson ImmunoResearch, West Grove, PA, USA).

Techniques: Expressing, Cell Culture, Phospho-proteomics, Activation Assay, Western Blot, Knockdown

Journal: International Journal of Oncology

Article Title: Expression and function of CXCL12/CXCR4/CXCR7 in thyroid cancer

doi: 10.3892/ijo.2016.3485

Figure Lengend Snippet: The expression of CXCL12, CXCR7 and CXCR4 in malignant and benign thyroid tissue specimens.

Article Snippet: The membrane was then incubated with the following primary antibodies: rabbit anti-human CXCR4, CXCR7, p-Akt1 (B-1), p-ERK (B-5), β-actin, GAPDH and ERK1/2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) at room temperature for 1 h or at 4°C overnight.

Techniques: Expressing

Journal: International Journal of Oncology

Article Title: Expression and function of CXCL12/CXCR4/CXCR7 in thyroid cancer

doi: 10.3892/ijo.2016.3485

Figure Lengend Snippet: Immunohistochemical staining of CXCR4 of thyroid tissue specimens. (A) Images of immunohistochemical staining with anti-CXCR4 for tissue specimens of papillary thyroid carcinoma, anaplastic thyroid carcinoma, Hashimoto's thyroiditis and nodular goiter. (B) Stronger CXCR4 staining in metastatic lesions than in the primary lesions.

Article Snippet: The membrane was then incubated with the following primary antibodies: rabbit anti-human CXCR4, CXCR7, p-Akt1 (B-1), p-ERK (B-5), β-actin, GAPDH and ERK1/2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) at room temperature for 1 h or at 4°C overnight.

Techniques: Immunohistochemical staining, Staining

Journal: International Journal of Oncology

Article Title: Expression and function of CXCL12/CXCR4/CXCR7 in thyroid cancer

doi: 10.3892/ijo.2016.3485

Figure Lengend Snippet: The intensity of CXCL12, CXCR7 and CXCR4 staining in thyroid tissue specimens.

Article Snippet: The membrane was then incubated with the following primary antibodies: rabbit anti-human CXCR4, CXCR7, p-Akt1 (B-1), p-ERK (B-5), β-actin, GAPDH and ERK1/2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) at room temperature for 1 h or at 4°C overnight.

Techniques: Staining

Journal: International Journal of Oncology

Article Title: Expression and function of CXCL12/CXCR4/CXCR7 in thyroid cancer

doi: 10.3892/ijo.2016.3485

Figure Lengend Snippet: Overexpression of CXCR4 and CXCR7 in K1 cells. (A) Representative image of western blot analysis of CXCR4 expression in K1 parental, mock-transfected, CXCR4-overexpression cells. (B) Representative image of western blot analysis of CXCR7 K1 parental, mock-transfected, CXCR7-overexpressing cells. Cells were harvested 72 h after transduction with CXCR4 or CXCR7 recombinant lentivirus, and protein extract was separated on SDS-PAGE gel and transferred to PVDF membrane. CXCR4 and CXCR7 protein were detected by anti-CXCR4 and anti-CXCR7. Densitometry was analyzed by Quantity One system.

Article Snippet: The membrane was then incubated with the following primary antibodies: rabbit anti-human CXCR4, CXCR7, p-Akt1 (B-1), p-ERK (B-5), β-actin, GAPDH and ERK1/2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) at room temperature for 1 h or at 4°C overnight.

Techniques: Over Expression, Western Blot, Expressing, Transfection, Transduction, Recombinant, SDS Page, Membrane

Journal: International Journal of Oncology

Article Title: Expression and function of CXCL12/CXCR4/CXCR7 in thyroid cancer

doi: 10.3892/ijo.2016.3485

Figure Lengend Snippet: Overexpression of CXCR4 and CXCR7 induces K1 cell invasion but has no effect on cell proliferation. (A) Overexpression of CXCR4 increased CXCL12-induced K1 cell invasion. Cells were treated with 100 ng/ml CXCL12 for 24 h. The concentration of control IgG and anti-CXCR4 was 10 μg/ml. The number of cells invaded through the membrane was counted 24 h after incubation with CXCL12. Three to 5 fields were randomly selected on each membrane and the average number of cells was used. The experiment was repeated at least 3 times. (B) Overexpression of CXCR4 or CXCR7 did not affect K1 cell proliferation. Cells were incubated in media containing 10% FBS, and cell growth was determined by using CCK-8 kit.

Article Snippet: The membrane was then incubated with the following primary antibodies: rabbit anti-human CXCR4, CXCR7, p-Akt1 (B-1), p-ERK (B-5), β-actin, GAPDH and ERK1/2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) at room temperature for 1 h or at 4°C overnight.

Techniques: Over Expression, Concentration Assay, Control, Membrane, Incubation, CCK-8 Assay

Journal: International Journal of Oncology

Article Title: Expression and function of CXCL12/CXCR4/CXCR7 in thyroid cancer

doi: 10.3892/ijo.2016.3485

Figure Lengend Snippet: The effect of overexpression of CXCR4 or CXCR7 on AKT and ERK phosphorylation and MMP-2 activity. (A) Overexpression of CXCR4 or CXCR7 increased AKT and ERK phosphorylation. Cells were harvested 72 h after transduction with CXCR4 or CXCR7 recombinant lentivirus, and protein extract was separated on SDS-PAGE gel and transferred to PVDF membrane. Densitometry was analyzed by Quantity One system. * P<0.05, significant difference between K1-CXCR7 and K1 mock. # P<0.05, significant difference between K1-CXCR4 and K1 mock. (B) Zymography assay of MMP activity. Culture media were collected and loaded in SDS-PAGE gel containing 1 mg/ml gelatin. After electrophoresis, the gel was incubated with MMP reaction buffer. The gel was then stained and de-stained. The experiment was repeated 3 times. Representative gel image is presented.

Article Snippet: The membrane was then incubated with the following primary antibodies: rabbit anti-human CXCR4, CXCR7, p-Akt1 (B-1), p-ERK (B-5), β-actin, GAPDH and ERK1/2 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) at room temperature for 1 h or at 4°C overnight.

Techniques: Over Expression, Phospho-proteomics, Activity Assay, Transduction, Recombinant, SDS Page, Membrane, Zymography, Electrophoresis, Incubation, Staining

Journal:

Article Title: The IL-8 regulated Chemokine Receptor CXCR7 Stimulates EGFR Signaling to Promote Prostate Cancer Growth

doi: 10.1158/0008-5472.CAN-10-2769

Figure Lengend Snippet: Expression of CXCR Chemokines and their receptors in normal prostate and tumor cells Total RNA isolated from various cultures was reverse transcribed and the cDNAs were quantified using q-PCR. Values shown are Fold differences (FD) as defined in the methods, normalized to GAPDH mRNA. ND: Not detected. Mean ± SD, n= 3

Article Snippet: In addition, we performed confocal microscopy of LNCaP cells labeled with anti-EGFR (mouse anti-human IgG) and anti-CXCR7 (Rabbit anti-human IgG, GeneTex) antibodies followed by, fluorescent labeling with secondary antibodies labeled with Alexafluor 555 and Alexfluor488, respectively.

Techniques: Expressing, Isolation

Journal:

Article Title: The IL-8 regulated Chemokine Receptor CXCR7 Stimulates EGFR Signaling to Promote Prostate Cancer Growth

doi: 10.1158/0008-5472.CAN-10-2769

Figure Lengend Snippet: A. Detection of CXCR4 and CXCR7 proteins in CaP and normal cells by Western blotting (LN: LNCaP; LA: LAPC-4; RW: RWPE-1). B. CXCR7 (i) and CXCR4 (ii) mRNA. C. CXCR7 protein levels in LNCaP and LAPC-4 cells continuously-expressing IL-8 (IL8-S) and their vector-only transfectants (e.g., IL-8V). D. Increased expression of CXCR7 mRNA (i) and protein levels (ii & iii) following IL-8 addition to growth factor-starved (24 h) RWPE-1 and LNCaP cells, respectively.

Article Snippet: In addition, we performed confocal microscopy of LNCaP cells labeled with anti-EGFR (mouse anti-human IgG) and anti-CXCR7 (Rabbit anti-human IgG, GeneTex) antibodies followed by, fluorescent labeling with secondary antibodies labeled with Alexafluor 555 and Alexfluor488, respectively.

Techniques: Western Blot, Expressing, Plasmid Preparation

Journal:

Article Title: The IL-8 regulated Chemokine Receptor CXCR7 Stimulates EGFR Signaling to Promote Prostate Cancer Growth

doi: 10.1158/0008-5472.CAN-10-2769

Figure Lengend Snippet: A, Decrease in cell density (cells/well, mean ± SEM) of CaP cell cultures following depletion of CXCR7 by siRNA. B (i)–ii), Decreased cell growth in PC-3 or LNCaP cells, stably-expressing either control-shRNA (V), or CXCR7 shRNA (T-74), determined by cell counting. Western blot analysis of CXCR7 in the cells transfected with V and T74 shRNA (Inset, i–ii). C, Cell density of PC-3 cultures expressing either control-shRNA (PC-3V), or CXCR7 shRNA (PC3T-74) in the presence or absence of CXCL11 (CX11) or SDF1 (10 ng /ml), cultured with 10% FBS or without FBS (ITS) medium for 5 days. D, Cell viability determined by MTT reduction assay in PC-3 cells transiently transfected with indicated siRNA. Data presented are mean ± S.E.M. n=3. *P < 0.05 versus control, Student's t test.

Article Snippet: In addition, we performed confocal microscopy of LNCaP cells labeled with anti-EGFR (mouse anti-human IgG) and anti-CXCR7 (Rabbit anti-human IgG, GeneTex) antibodies followed by, fluorescent labeling with secondary antibodies labeled with Alexafluor 555 and Alexfluor488, respectively.

Techniques: Stable Transfection, Expressing, shRNA, Cell Counting, Western Blot, Transfection, Cell Culture, MTT Reduction Assay

Journal:

Article Title: The IL-8 regulated Chemokine Receptor CXCR7 Stimulates EGFR Signaling to Promote Prostate Cancer Growth

doi: 10.1158/0008-5472.CAN-10-2769

Figure Lengend Snippet: A. Percent of cells in each cell-cycle phase was estimated from flow cytometry analysis of DNA contents in CaP cells 48 hours after transfection with CXCR7 siRNA or c-siRNA. B. DNA content histograms of CaP cells transfected with c-siRNA or CXCR7 siRNA. C, Comparison of expression of cell cycle regulatory proteins by Western blotting as in A, and relative expression (R.I.) shown are normalized to protein band intensities of β-actin. D, Levels of P- Erk1/2 measured with an ELISA in C-siRNA (c-si), CXCR7 siRNA (CXC7si), CXCR7shRNA (T74) or V transfectants following EGF stimulation (5 nM, 10 min). The p-Erk1/2 levels were lower in CXCR7 depleted PC-3 cells (27% in siRNA and 63% in shRNA depleted PC-3-cells) (* = P ≤0.05, n=3).

Article Snippet: In addition, we performed confocal microscopy of LNCaP cells labeled with anti-EGFR (mouse anti-human IgG) and anti-CXCR7 (Rabbit anti-human IgG, GeneTex) antibodies followed by, fluorescent labeling with secondary antibodies labeled with Alexafluor 555 and Alexfluor488, respectively.

Techniques: Flow Cytometry, Transfection, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, shRNA

Journal:

Article Title: The IL-8 regulated Chemokine Receptor CXCR7 Stimulates EGFR Signaling to Promote Prostate Cancer Growth

doi: 10.1158/0008-5472.CAN-10-2769

Figure Lengend Snippet: A, Increase in cell density of RWPE-1 cultures stably expressing CXCR7 (RWCX7) as compared to RW-V cells, B, A comparison of levels of p-EGFR, p-Erk1/2 and cyclinD1 in cells described in A and CaP cells. C, Levels of p-EGFR and p-Erk1/2 in CaP cells following transient transfection with either c-siRNA (c-si) or CXCR7 siRNA (CX7) and stimulated with EGF (5 nM) for 5 min. CXCR7siRNA transfectants had lower level of p-EGFR and p-Erk1/2 compared to that of c-siRNA transfected cells (C Lanes 2 and 4). D, Western blots of pEGFRy1110 from cells described in A that were growth factor starved for 24 h and then stimulated with 5 nM EGF, CXCL11 or SDF-1 (both 100 ng/ml). Total EGFR and ß-actin levels are shown as loading controls.

Article Snippet: In addition, we performed confocal microscopy of LNCaP cells labeled with anti-EGFR (mouse anti-human IgG) and anti-CXCR7 (Rabbit anti-human IgG, GeneTex) antibodies followed by, fluorescent labeling with secondary antibodies labeled with Alexafluor 555 and Alexfluor488, respectively.

Techniques: Stable Transfection, Expressing, Transfection, Western Blot

Journal:

Article Title: The IL-8 regulated Chemokine Receptor CXCR7 Stimulates EGFR Signaling to Promote Prostate Cancer Growth

doi: 10.1158/0008-5472.CAN-10-2769

Figure Lengend Snippet: A–B, Demonstration of coimmunoprecipitation of CXCR7 and EGFR. Cell lysates of PC-3 and LNCaP were immunoprecipitated with CXCR7 antibody followed by western blotting against p-EGFR and vice versa. C, EGFR co-precipitated with CXCR7 in RWCX7 cells. RWCX7 cells or control vector (RW-V), were immunoprecipitated with CXCR7 antibody followed by western blotting with an anti- p-EGFR (pY1110) or an antibody recognizing phospho-Ser at residue 1070-71 in EGFR (pS1070-71) (Epitomics).

Article Snippet: In addition, we performed confocal microscopy of LNCaP cells labeled with anti-EGFR (mouse anti-human IgG) and anti-CXCR7 (Rabbit anti-human IgG, GeneTex) antibodies followed by, fluorescent labeling with secondary antibodies labeled with Alexafluor 555 and Alexfluor488, respectively.

Techniques: Immunoprecipitation, Western Blot, Plasmid Preparation

Journal:

Article Title: The IL-8 regulated Chemokine Receptor CXCR7 Stimulates EGFR Signaling to Promote Prostate Cancer Growth

doi: 10.1158/0008-5472.CAN-10-2769

Figure Lengend Snippet: A, Tumor growth over time in athymic mice was recorded for mice injected with PC-3 cells stably expressing CXCR7 shRNA (T73 and T74) or scrambled sequence-shRNA (V) B, Blots of CXCR7, cyclin D1 p-EGFR CXCR7 tumor tissue of PC-3V, T73 and T74 C–D, CXCR7 and VEGF mRNA level were decreased in T73 and T74 tumors as analyzed by q-PCR. There was a significant delay (p<0.03) in tumor growth and in the terminal tumor volume of T73 and T74 tumors.

Article Snippet: In addition, we performed confocal microscopy of LNCaP cells labeled with anti-EGFR (mouse anti-human IgG) and anti-CXCR7 (Rabbit anti-human IgG, GeneTex) antibodies followed by, fluorescent labeling with secondary antibodies labeled with Alexafluor 555 and Alexfluor488, respectively.

Techniques: Injection, Stable Transfection, Expressing, shRNA, Sequencing

Journal: Experimental and Therapeutic Medicine

Article Title: Down-regulation of CXCR7 inhibits the growth and lung metastasis of human hepatocellular carcinoma cells with highly metastatic potential

doi: 10.3892/etm.2011.358

Figure Lengend Snippet: Table I.

Article Snippet: Rabbit anti-human CXCR7 IgG at 1:200 and rabbit anti-human CXCR4 IgG (Santa Cruz Biotechnology) at 1:100 were used as the primary antibodies for detection.

Techniques: In Vivo