Journal: The Journal of Experimental Medicine

Article Title: Superior antigen cross-presentation and XCR1 expression define human CD11c + CD141 + cells as homologues of mouse CD8 + dendritic cells

doi: 10.1084/jem.20100348

Figure Lengend Snippet: XCL1 induces a [Ca 2+ ] i signal in CD141 + DCs. CD141 + DCs were flow sorted to a purity >98.7%, immobilized on poly– l -lysine–coated glass coverslips, and loaded with 2 µM fura-2/AM. Cells were imaged in a monochromator-assisted digital video imaging system and challenged with 1 µg/ml XCL1 as indicated (left arrow). Subsequently, the same cells were challenged again with a mixture of 100 ng/ml CCL2, 200 ng/ml CCL21, 200 ng/ml CXCL9, and 1 ng/ml CX3CL1 used as a positive control (right arrow). The data shown represent [Ca 2+ ] i concentrations of 300 single cells (gray lines) measured in two independent experiments. The mean [Ca 2+ ] i signal averaged over all cells responding to XCL1 is indicated (black line).

Article Snippet: The lower chamber was filled with chemotaxis medium containing recombinant human XCL1 (R&D Systems) or any of the chemokines CCL2 (100 ng/ml), CCL21 (200 ng/ml), CX3CL1 (1 ng/ml), CXCL12 (200 ng/ml for T cells, B cells, NK cells, and monocytes; 100 ng/ml for pDCs), and CXCL8 (100 ng/ml; all from R&D Systems), and the cells were incubated for 150 min at 37°C in 5% CO 2 .

Techniques: Imaging, Positive Control

Journal: The Journal of Experimental Medicine

Article Title: Superior antigen cross-presentation and XCR1 expression define human CD11c + CD141 + cells as homologues of mouse CD8 + dendritic cells

doi: 10.1084/jem.20100348

Figure Lengend Snippet: XCL1 selectively induces chemotaxis in CD141 + DCs. (A) A mixture of highly purified, flow-sorted DC subtypes (20% CD141 + , 40% CD16 + , and 40% CD1c + DCs; Input DC) was tested for migration in response to medium alone or to serial dilutions of XCL1 (10–5,000 ng/ml) in a Transwell system. A combination of the chemokines CCL2, CCL21, and CX3CL1 was used as a positive control for the DC subsets (Migrated DC). The absolute numbers of CD141 + , CD1c + , and CD16 + DCs in input and migrated cell populations are truly represented in the dot plots, because all cells within a defined volume were included in the analysis in each instance. (B) Proportion of migrated CD1c + , CD16 + , and CD141 + DCs in the experiment shown in A. (C) Proportion of migrated pDCs, monocytes, granulocytes, T cells, B cells, and NK cells in response to XCL1 (10–1,000 ng/ml) or the chemokines CXCL12 and CXCL8, which were used as positive controls. For migration assays of B cells, NK cells, and monocytes, PBMCs were magnetically depleted of T cells, and for T cell migration, PBMCs were used directly. For migration assays of granulocytes, whole blood cells were used after erythrocyte lysis with ACK buffer, and pDCs were magnetically enriched from PBMCs with the Plasmacytoid Dendritic Cell Isolation Kit (Miltenyi Biotec). All experiments with DCs were performed three times; all other populations were assayed twice. Error bars represent means ± SEM.

Article Snippet: The lower chamber was filled with chemotaxis medium containing recombinant human XCL1 (R&D Systems) or any of the chemokines CCL2 (100 ng/ml), CCL21 (200 ng/ml), CX3CL1 (1 ng/ml), CXCL12 (200 ng/ml for T cells, B cells, NK cells, and monocytes; 100 ng/ml for pDCs), and CXCL8 (100 ng/ml; all from R&D Systems), and the cells were incubated for 150 min at 37°C in 5% CO 2 .

Techniques: Chemotaxis Assay, Purification, Migration, Positive Control, Lysis, Cell Isolation

Journal: The Journal of Experimental Medicine

Article Title: Superior antigen cross-presentation and XCR1 expression define human CD11c + CD141 + cells as homologues of mouse CD8 + dendritic cells

doi: 10.1084/jem.20100348

Figure Lengend Snippet: Involvement of the XCL1–XCR1 communication axis in the innate and adaptive cytotoxic responses to cross-presented microbial and tumor antigens. Secretion of the chemokine XCL1 by activated NK cells specifically attracts XCR1-expressing DCs capable of antigen cross-presentation. This ensures an effective communication between these cells in the innate phase of the immune response. In the adaptive phase, secretion of XCL1 by activated CD8 + T cells optimizes the communication with antigen cross-presenting DCs and facilitates the differentiation of CD8 + T cells to cytotoxic cells.

Article Snippet: The lower chamber was filled with chemotaxis medium containing recombinant human XCL1 (R&D Systems) or any of the chemokines CCL2 (100 ng/ml), CCL21 (200 ng/ml), CX3CL1 (1 ng/ml), CXCL12 (200 ng/ml for T cells, B cells, NK cells, and monocytes; 100 ng/ml for pDCs), and CXCL8 (100 ng/ml; all from R&D Systems), and the cells were incubated for 150 min at 37°C in 5% CO 2 .

Techniques: Expressing

Journal: Oncology Reports

Article Title: Zinc finger protein 695 facilitates the proliferation of colorectal cancer cells through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways

doi: 10.3892/or.2025.8949

Figure Lengend Snippet: Clinical relevance of ZNF695 in patients with CRC. (A) Transcript levels of ZNF695 in pan-cancer was analyzed using UALCAN based on TCGA data. *P<0.05 vs. normal group. (B) Transcript levels of ZNF695 in cancer (n=275) and non-cancer (n=349) tissues of patients with CRC. *P<0.05. (C) mRNA expression of ZNF695 in CRC tissues based on the individual cancer stage and in non-cancer tissues was analyzed using Gene Expression Profiling Interactive Analysis. ***P<0.001. (D) Promoter methylation level of ZNF695 in cancer (n=313) and non-cancer (n=37) tissues of patients with CRC was analyzed using UALCAN. P>0.05. (E) Promoter methylation level of ZNF695 in in CRC tissues based on the individual cancer stage and in non-cancer tissues was analyzed using UALCAN. Normal vs. stage 2, P<0.05. (F) Overall survival of patients with CRC who were divided into the ZNF695 high expression group (n=406) and the low expression group (n=655) was analyzed from Kaplan Meier plotter. P=0.065. (G) mRNA expression levels of ZNF695 were detected by reverse transcription-quantitative PCR in NCM460, HT-29, HCT-116 and HCT-8 cells. *P<0.05, **P<0.01 vs. NCM460. BLCA, bladder urothelial carcinoma; BRCA, breast invasive carcinoma; CESC, cervical squamous cell carcinoma and endocervical adenocarcinoma; CHOL, cholangiocarcinoma; CRC, colorectal cancer; ESCA, esophageal carcinoma; GBM, glioblastoma; HNSC, head and neck squamous cell carcinoma; KICH, kidney chromophobe; KIRC, kidney renal clear cell carcinoma; KIRP, kidney renal papillary cell carcinoma; LIHC, liver hepatocellular carcinoma; LUAD, lung adenocarcinoma; LUSC, lung squamous cell carcinoma; PAAD, pancreatic adenocarcinoma; PRAD, prostate adenocarcinoma; PCPG, pheochromocytoma and paraganglioma; READ, rectum adenocarcinoma; SARC, sarcoma; SKCM, skin cutaneous melanoma; THCA, thyroid carcinoma; THYM, thymoma; STAD, stomach adenocarcinoma; UCEC, uterine corpus endometrial carcinoma; TCGA, The Cancer Genome Atlas; ZNF695, zinc finger protein 695.

Article Snippet: The following primary antibodies were used: ZNF695 (cat. no. 25556-1-AP; 1:800; Proteintech Group, Inc.), NEK2 (cat. no. 24171-1-AP; 1:800; Proteintech Group Inc.), Akt (cat. no. 9272; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (cat. no. 4060; 1:1,000; Cell Signaling Technology, Inc.), ribosomal protein S6 (S6; cat. no. 2217; 1:1,000; Cell Signaling Technology, Inc.), p-S6 (cat. no. 2211; 1:1,000; Cell Signaling Technology, Inc.), p21 (cat. no. 2947; 1:1,000; Cell Signaling Technology, Inc.), Bcl-2 (cat. no. 3498; 1:1,000; Cell Signaling Technology, Inc.), β-actin (cat. no. 66009-1-Ig; 1:5,000; Proteintech Group Inc.) and GAPDH (cat. no. 60004-1-Ig; 1:5,000; Proteintech Group Inc.).

Techniques: Expressing, Gene Expression, Methylation, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Oncology Reports

Article Title: Zinc finger protein 695 facilitates the proliferation of colorectal cancer cells through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways

doi: 10.3892/or.2025.8949

Figure Lengend Snippet: Overexpression of ZNF695 supports the proliferation of CRC cells. (A) mRNA and protein levels of ZNF695 in siCtrl, siZNF695#1 and siZNF695#2 HCT-8 cells were detected using RT-qPCR and western blotting, respectively. **P<0.01 vs. siCtrl group. (B) Proliferation of HCT-8 cells transfected with siCtrl, siZNF695#1 and siZNF695#2 was determined using a CCK8 assay. *P<0.05, **P<0.01 vs. siCtrl group. (C) mRNA and protein levels of ZNF695 in Ctrl and ZNF695-overexpressing HT-29 cells were detected using RT-qPCR and western blotting, respectively. **P<0.01 vs. Ctrl group. (D) Proliferation of Ctrl and ZNF695-overexpressing HT-29 cells was determined by CCK8 assay. *P<0.05, **P<0.01 vs. Ctrl group. Colony formation was assessed in (E) siCtrl, siZNF695#1 and siZNF695#2 HCT-8 cells, and in Ctrl and ZNF695-overexpressing HT-29 cells. (F) Quantification colony numbers *P<0.05, **P<0.01vs. siCtrl, ## P<0.01 vs. Ctrl group. CRC, colorectal cancer; RT-qPCR, reverse transcription-quantitative PCR; CCK8, Cell Counting Kit-8; ZNF695, zinc finger protein 695; si, small interfering.

Article Snippet: The following primary antibodies were used: ZNF695 (cat. no. 25556-1-AP; 1:800; Proteintech Group, Inc.), NEK2 (cat. no. 24171-1-AP; 1:800; Proteintech Group Inc.), Akt (cat. no. 9272; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (cat. no. 4060; 1:1,000; Cell Signaling Technology, Inc.), ribosomal protein S6 (S6; cat. no. 2217; 1:1,000; Cell Signaling Technology, Inc.), p-S6 (cat. no. 2211; 1:1,000; Cell Signaling Technology, Inc.), p21 (cat. no. 2947; 1:1,000; Cell Signaling Technology, Inc.), Bcl-2 (cat. no. 3498; 1:1,000; Cell Signaling Technology, Inc.), β-actin (cat. no. 66009-1-Ig; 1:5,000; Proteintech Group Inc.) and GAPDH (cat. no. 60004-1-Ig; 1:5,000; Proteintech Group Inc.).

Techniques: Over Expression, Quantitative RT-PCR, Western Blot, Transfection, CCK-8 Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Cell Counting

Journal: Oncology Reports

Article Title: Zinc finger protein 695 facilitates the proliferation of colorectal cancer cells through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways

doi: 10.3892/or.2025.8949

Figure Lengend Snippet: ZNF695 modulates the apoptosis and cell cycle progression of CRC cells. (A) Cell cycle progression of siCtrl, siZNF695#1 and siZNF695#2 HCT-8 cells was measured by staining the cells with PI and via flow cytometric analysis. (B) Cell cycle progression of Ctrl and ZNF695-overexpressing HT-29 cells was detected by staining the cells with PI and via flow cytometric analysis. (C) Quantification of cell cycle results as shown in A and B. *P<0.05 vs. siCtrl, # P<0.05 vs. Ctrl group. (D) Apoptosis of siCtrl, siZNF695#1 and siZNF695#2 HCT-8 cells was assessed by staining the cells with Annexin V and PI, followed by flow cytometric analysis. (E) Apoptosis of Ctrl and ZNF695-overexpressing HT-29 cells was assessed by staining the cells with Annexin V and PI, followed by flow cytometric analysis. (F) Quantification of apoptosis results as shown in D and E. *P<0.05 vs. siCtrl, # P<0.05 vs. Ctrl group. (G) Caspase 3/7 activity was measured in HCT-8 cells transfected with siCtrl, siZNF695#1 and siZNF695#2 and in HT-29 cells infected with Ctrl and ZNF695-overexpressing lentiviruses. *P<0.05 vs. siCtrl, # P<0.05 vs. Ctrl group. CRC, colorectal cancer; ZNF695, zinc finger protein 695; si, small interfering.

Article Snippet: The following primary antibodies were used: ZNF695 (cat. no. 25556-1-AP; 1:800; Proteintech Group, Inc.), NEK2 (cat. no. 24171-1-AP; 1:800; Proteintech Group Inc.), Akt (cat. no. 9272; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (cat. no. 4060; 1:1,000; Cell Signaling Technology, Inc.), ribosomal protein S6 (S6; cat. no. 2217; 1:1,000; Cell Signaling Technology, Inc.), p-S6 (cat. no. 2211; 1:1,000; Cell Signaling Technology, Inc.), p21 (cat. no. 2947; 1:1,000; Cell Signaling Technology, Inc.), Bcl-2 (cat. no. 3498; 1:1,000; Cell Signaling Technology, Inc.), β-actin (cat. no. 66009-1-Ig; 1:5,000; Proteintech Group Inc.) and GAPDH (cat. no. 60004-1-Ig; 1:5,000; Proteintech Group Inc.).

Techniques: Staining, Activity Assay, Transfection, Infection

Journal: Oncology Reports

Article Title: Zinc finger protein 695 facilitates the proliferation of colorectal cancer cells through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways

doi: 10.3892/or.2025.8949

Figure Lengend Snippet: ZNF695 affects the expression of p21 and Bcl-2. (A) Western blot analysis of p21, Bcl-2 and GAPDH in (A) HCT-8 cells transfected with siCtrl, siZNF695#1 and siZNF695#2, and in (B) HT-29 cells infected with Ctrl and ZNF695-overexpressing lentiviruses. GAPDH served as internal control. ZNF695, zinc finger protein 695; si, small interfering.

Article Snippet: The following primary antibodies were used: ZNF695 (cat. no. 25556-1-AP; 1:800; Proteintech Group, Inc.), NEK2 (cat. no. 24171-1-AP; 1:800; Proteintech Group Inc.), Akt (cat. no. 9272; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (cat. no. 4060; 1:1,000; Cell Signaling Technology, Inc.), ribosomal protein S6 (S6; cat. no. 2217; 1:1,000; Cell Signaling Technology, Inc.), p-S6 (cat. no. 2211; 1:1,000; Cell Signaling Technology, Inc.), p21 (cat. no. 2947; 1:1,000; Cell Signaling Technology, Inc.), Bcl-2 (cat. no. 3498; 1:1,000; Cell Signaling Technology, Inc.), β-actin (cat. no. 66009-1-Ig; 1:5,000; Proteintech Group Inc.) and GAPDH (cat. no. 60004-1-Ig; 1:5,000; Proteintech Group Inc.).

Techniques: Expressing, Western Blot, Transfection, Infection, Control

Journal: Oncology Reports

Article Title: Zinc finger protein 695 facilitates the proliferation of colorectal cancer cells through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways

doi: 10.3892/or.2025.8949

Figure Lengend Snippet: ZNF695 positively regulates the expression of NEK2 in CRC cells. (A) List of genes that were positively correlated with ZNF695 in patients with CRC based on The Cancer Genome Atlas database. ***P<0.001. mRNA expression levels of NEK2 were analyzed by reverse transcription-qPCR in (B) HCT-8 cells transfected with siCtrl, siZNF695#1 and siZNF695#2, and in (C) HT-29 cells infected with Ctrl and ZNF695-overexpressing lentiviruses. *P<0.05 vs. siCtrl group; **P<0.01 vs. Ctrl group. Dual luciferase activity of the NEK2 gene promoter was detected in (D) HCT-8 cells transfected with siCtrl, siZNF695#1 and siZNF695#2, and in (E) HT-29 cells infected with Ctrl and ZNF695-overexpressing lentiviruses. *P<0.05 vs. siCtrl group; **P<0.01 vs. Ctrl group. (F) HT-29 cells were transfected with ZNF695-Flag-pCDNA3.1 vectors, and the cells were subjected to ChIP-qPCR experiments to assess whether ZNF695 interacted with the promoter sequence of the NEK2 gene. Based on The Cancer Genome Atlas database of patients with CRC. **P<0.01 vs. IgG group. (G) there was a positive correlation between ZNF695 and NEK2 (R=0.62; P<0.0001), and (H) a significant difference between the transcript levels of NEK2 in cancer (n=275) and non-cancer (n=349) tissues. *P<0.05. CRC, colorectal cancer; qPCR, quantitative PCR; ChIP, chromatin immunoprecipitation; ZNF695, zinc finger protein 695; si, small interfering; NEK2, NIMA-related kinase 2.

Article Snippet: The following primary antibodies were used: ZNF695 (cat. no. 25556-1-AP; 1:800; Proteintech Group, Inc.), NEK2 (cat. no. 24171-1-AP; 1:800; Proteintech Group Inc.), Akt (cat. no. 9272; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (cat. no. 4060; 1:1,000; Cell Signaling Technology, Inc.), ribosomal protein S6 (S6; cat. no. 2217; 1:1,000; Cell Signaling Technology, Inc.), p-S6 (cat. no. 2211; 1:1,000; Cell Signaling Technology, Inc.), p21 (cat. no. 2947; 1:1,000; Cell Signaling Technology, Inc.), Bcl-2 (cat. no. 3498; 1:1,000; Cell Signaling Technology, Inc.), β-actin (cat. no. 66009-1-Ig; 1:5,000; Proteintech Group Inc.) and GAPDH (cat. no. 60004-1-Ig; 1:5,000; Proteintech Group Inc.).

Techniques: Expressing, Reverse Transcription, Transfection, Infection, Luciferase, Activity Assay, ChIP-qPCR, Sequencing, Real-time Polymerase Chain Reaction, Chromatin Immunoprecipitation

Journal: Oncology Reports

Article Title: Zinc finger protein 695 facilitates the proliferation of colorectal cancer cells through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways

doi: 10.3892/or.2025.8949

Figure Lengend Snippet: Overexpression of ZNF695 activates the Akt/mTOR signaling pathway and dictates the sensitivity of CRC cells to rapamycin. (A) Western blot analysis of NEK2, p-Akt, Akt, p-S6, S6 and GAPDH in HCT-8 cells transfected with siCtrl, siZNF695#1 and siZNF695#2. GAPDH served as an internal control. (B) HCT-8 cells transfected with siCtrl, siZNF695#1 and siZNF695#2 were treated with different concentrations of rapamycin for 48 h and cell viability was determined by CCK8 assay. *P<0.05, **P<0.01 vs. siCtrl group. (C) Growth inhibition rate in response to different concentrations of rapamycin was calculated in HCT-8 cells transfected with siCtrl, siZNF695#1 and siZNF695#2. *P<0.05 vs. siCtrl group. (D) Western blot analysis of NEK2, p-Akt, Akt, p-S6, S6 and GAPDH in HT-29 cells infected with Ctrl and ZNF695-overexpressing lentiviruses. GAPDH served as an internal control. (E) HT-29 cells infected with Ctrl and ZNF695-overexpressing lentiviruses were treated with different concentrations of rapamycin for 48 h and cell viability was determined by CCK8 assay. *P<0.05, **P<0.01 vs. Ctrl group. (F) Growth inhibition rate in response to different concentrations of rapamycin was calculated in HT-29 cells infected with Ctrl and ZNF695-overexpressing lentiviruses. *P<0.05 vs. Ctrl group. CRC, colorectal cancer; CCK8, Cell Counting Kit-8; ZNF695, zinc finger protein 695; si, small interfering; p-, phosphorylated; NEK2, NIMA-related kinase 2; S6, ribosomal protein S6.

Article Snippet: The following primary antibodies were used: ZNF695 (cat. no. 25556-1-AP; 1:800; Proteintech Group, Inc.), NEK2 (cat. no. 24171-1-AP; 1:800; Proteintech Group Inc.), Akt (cat. no. 9272; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (cat. no. 4060; 1:1,000; Cell Signaling Technology, Inc.), ribosomal protein S6 (S6; cat. no. 2217; 1:1,000; Cell Signaling Technology, Inc.), p-S6 (cat. no. 2211; 1:1,000; Cell Signaling Technology, Inc.), p21 (cat. no. 2947; 1:1,000; Cell Signaling Technology, Inc.), Bcl-2 (cat. no. 3498; 1:1,000; Cell Signaling Technology, Inc.), β-actin (cat. no. 66009-1-Ig; 1:5,000; Proteintech Group Inc.) and GAPDH (cat. no. 60004-1-Ig; 1:5,000; Proteintech Group Inc.).

Techniques: Over Expression, Western Blot, Transfection, Control, CCK-8 Assay, Inhibition, Infection, Cell Counting

Journal: Oncology Reports

Article Title: Zinc finger protein 695 facilitates the proliferation of colorectal cancer cells through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways

doi: 10.3892/or.2025.8949

Figure Lengend Snippet: Knockdown of NEK2 rescues the tumor-promoting function of ZNF695. (A) Protein levels of NEK2 in Ctrl, ZNF695 and ZNF695 + siNEK2 HT-29 cells were detected by western blotting. (B) Proliferation of Ctrl, ZNF695 and ZNF695 + siNEK2 HT-29 cells was determined by Cell Counting Kit-8 assay. *P<0.05, **P<0.01 vs. Ctrl, # P<0.05, ## P<0.01 vs. ZNF695. (C) Colony formation was assess in Ctrl, ZNF695 and ZNF695 + siNEK2 HT-29 cells. *P<0.05 (Ctrl vs. ZNF695), # P<0.05 (ZNF695 vs. ZNF695 + siNEK2). (D) Caspase 3/7 activity was measured in Ctrl, ZNF695 and ZNF695 + siNEK2 HT-29 cells. *P<0.05, (Ctrl vs. ZNF695), # P<0.05 (ZNF695 vs. ZNF695 + siNEK2). CRC, colorectal cancer; ZNF695, zinc finger protein 695; si, small interfering; NEK2, NIMA-related kinase 2.

Article Snippet: The following primary antibodies were used: ZNF695 (cat. no. 25556-1-AP; 1:800; Proteintech Group, Inc.), NEK2 (cat. no. 24171-1-AP; 1:800; Proteintech Group Inc.), Akt (cat. no. 9272; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (cat. no. 4060; 1:1,000; Cell Signaling Technology, Inc.), ribosomal protein S6 (S6; cat. no. 2217; 1:1,000; Cell Signaling Technology, Inc.), p-S6 (cat. no. 2211; 1:1,000; Cell Signaling Technology, Inc.), p21 (cat. no. 2947; 1:1,000; Cell Signaling Technology, Inc.), Bcl-2 (cat. no. 3498; 1:1,000; Cell Signaling Technology, Inc.), β-actin (cat. no. 66009-1-Ig; 1:5,000; Proteintech Group Inc.) and GAPDH (cat. no. 60004-1-Ig; 1:5,000; Proteintech Group Inc.).

Techniques: Knockdown, Western Blot, Cell Counting, Activity Assay

Journal: Oncology Reports

Article Title: Zinc finger protein 695 facilitates the proliferation of colorectal cancer cells through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways

doi: 10.3892/or.2025.8949

Figure Lengend Snippet: Diagram of the ZNF695 signaling pathway in colorectal cancer tumorigenesis. ZNF695, zinc finger protein 695; NEK2, NIMA-related kinase 2.

Article Snippet: The following primary antibodies were used: ZNF695 (cat. no. 25556-1-AP; 1:800; Proteintech Group, Inc.), NEK2 (cat. no. 24171-1-AP; 1:800; Proteintech Group Inc.), Akt (cat. no. 9272; 1:1,000; Cell Signaling Technology, Inc.), phosphorylated (p)-Akt (cat. no. 4060; 1:1,000; Cell Signaling Technology, Inc.), ribosomal protein S6 (S6; cat. no. 2217; 1:1,000; Cell Signaling Technology, Inc.), p-S6 (cat. no. 2211; 1:1,000; Cell Signaling Technology, Inc.), p21 (cat. no. 2947; 1:1,000; Cell Signaling Technology, Inc.), Bcl-2 (cat. no. 3498; 1:1,000; Cell Signaling Technology, Inc.), β-actin (cat. no. 66009-1-Ig; 1:5,000; Proteintech Group Inc.) and GAPDH (cat. no. 60004-1-Ig; 1:5,000; Proteintech Group Inc.).

Techniques:

Journal: The Journal of rheumatology

Article Title: Expression and function of the C-class chemokine lymphotactin (XCL1) in Wegener's granulomatosis.

doi: 10.3899/jrheum.090244

Figure Lengend Snippet: Figure 2. Flow cytometric analysis results of XCL1 expression. PBMC isolated from WG patients and healthy controls were subjected to flow cytometric analysis after stimulation with PMA/ionomycin. XCL1 was expressed in a significantly greater proportion of CD4+ and CD8+ T cells in WG patients in comparison to controls (A). XCL1 was detected in WG patients in a significantly greater proportion of CD4+ T cells lacking the costimulatory molecule CD28 (p = 0.007) (B). In CD8+ T cells, XCL1 expression was found mainly within the CD8+CD28– T cell subpopulation in WG patients, but there was no statistically significant difference of frequencies of XCL1-expressing CD8+CD28– T cells between WG patients and controls (C). Comparing T cells from patients with active and inactive WG, a significant difference was detected between these patient groups in frequencies of XCL1-positive CD4+ T cells and CD8+ T cells (p < 0.001) (D).

Article Snippet: Cells were subsequently stimulated with human recombinant XCL1 (R&D Systems, Wiesbaden, Germany) at a concentration of 100 ng/ml for 24 h. To control the specificity of XCL1 action, anti-human XCL1 antibody (R&D Systems) was added to the control wells.

Techniques: Expressing, Isolation, Comparison