Journal: American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons

Article Title: Implanted adipose-derived stem cells attenuate small-for-size liver graft injury by secretion of VEGF in rats.

doi: 10.1111/j.1600-6143.2011.03870.x

Figure Lengend Snippet: Figure 1: Identification and purity of ADSCs. Flow cytometry revealed that the distribution of ADSCs that stained for CD14, CD31, CD34, CD45, CD68 and CD133 (shaded regions) did not differ from that of the isotype control (open regions). The majority of cells positively stained for CD29 and CD90 (shaded regions) compared with the isotype control cells (open regions) (A). Immunofluorescence analysis confirmed that ADSCs stained positively for CD29 and CD90, but not CD14, CD31, CD34, CD45, CD68 and CD133 (B). ADSCs resembled fibroblasts in culture and were successfully induced into adipocytes and osteocytes (C).

Article Snippet: Passage 3 cells were verified by immunostaining of surface markers for analysis by flow cytometry (Beckman Coulter, Brea, CA, USA) and immunofluorescence microscopy (Olympus, Tokyo, Japan) using fluorescent-conjugated antibodies against rat CD14, CD34, CD45, CD90 (Caltag Laboratories, San Francisco, CA, USA), CD29 (Biolegend, San Diego, CA, USA), CD31, CD68 (Abcam, Cambridge, MA, USA), CD133 (Novus Biologicals, Littleton, CO, USA) and respective isotypes.

Techniques: Flow Cytometry, Staining, Control

Journal: OncoTargets and therapy

Article Title: NFATC2 is a novel therapeutic target for colorectal cancer stem cells

doi: 10.2147/OTT.S169129

Figure Lengend Snippet: NFATC2 is upregulated in CRC-SCs. ( A ) qRT-PCR analysis of NFATC2 in primary CRC spheres and re-adherent cells relative to adherent cells. Primary CRC cells were isolated from the cancer tissues of CRC patients (No 1, 2, 3, and 4). Spheres were obtained by suspension culture. ( B ) Western blot analysis of NFATC2 in primary CRC spheres and adherent and re-adherent cells. ( C ) qRT-PCR analysis of NFATC2 in sorted CD44 + (left) or CD133 + (right) primary CRC cells relative to negative cells. Primary CRC cells were isolated from the cancer tissues of colorectal cancer patients (No 1, 2, 3, and 4). CD44 + or CD133 + cells were obtained by flow cytometry. ( D ) The correlation between the transcription level of NFATC2 and CD44 (left) and CD133 (right) in pin primary CRC sphere-derived cells. The mRNA level of each gene was determined by qRT-PCR. Data were normalized to GAPDH as ∆CT and analyzed by Spearman’s correlation analysis. Data are represented as mean ± SD; * P <0.05, *** P <0.001; two-tailed Student’s t -test. Abbreviations: CRC, colorectal cancer; NFATC2, nuclear factor of activated T-cells, cytoplasmic 2; CRC-SCs, colorectal cancer stem cells; qRT-PCR, quantitative real-time polymerase chain reaction.

Article Snippet: CD133 , IB , Mouse monoclonal , , Miltenyi Biotec , 130-092-395 , 100.

Techniques: Quantitative RT-PCR, Isolation, Suspension, Western Blot, Flow Cytometry, Derivative Assay, Two Tailed Test, Real-time Polymerase Chain Reaction

Journal: OncoTargets and therapy

Article Title: NFATC2 is a novel therapeutic target for colorectal cancer stem cells

doi: 10.2147/OTT.S169129

Figure Lengend Snippet: Overexpression of NFATC2 promotes the stemness of CRC cells. ( A , B ) Sphere formation assay of NFATC2-overexpressing and control primary CRC cells. Primary CRC cells were isolated from the cancer tissues of CRC patients (No 1 and 2). The 1st passaged spheres were obtained by suspension culture for 15 days and the number and average diameters of the spheres were counted ( A ). The number of 1st, 2nd, and 3rd passaged spheres isolated from the cancer tissues of CRC patients (No 1, 2, 3, and 4) was also counted ( B ). ( C ) qRT-PCR analysis of CD44 (left) and CD133 (right) in NFATC2-overexpressing and control primary CRC cells. Primary CRC cells were isolated from the cancer tissues of CRC patients (No 1 and 2). NFATC2-overexpressing and control cells were generated by lentivirus delivery system. ( D ) Western blot analysis of CD44 and CD133 in NFATC2-overexpressing and control primary CRC cells. Data are represented as mean ± SD; *** P <0.001; two-tailed Student’s t -test. Abbreviations: CRC, colorectal cancer; NFATC2, nuclear factor of activated T-cells, cytoplasmic 2; CRC-SCs, colorectal cancer stem cells; qRT-PCR, quantitative real-time polymerase chain reaction.

Article Snippet: CD133 , IB , Mouse monoclonal , , Miltenyi Biotec , 130-092-395 , 100.

Techniques: Over Expression, Tube Formation Assay, Control, Isolation, Suspension, Quantitative RT-PCR, Generated, Western Blot, Two Tailed Test, Real-time Polymerase Chain Reaction

Journal: OncoTargets and therapy

Article Title: NFATC2 is a novel therapeutic target for colorectal cancer stem cells

doi: 10.2147/OTT.S169129

Figure Lengend Snippet: Knockdown of NFATC2 inhibits the stemness of CRC cells. ( A , B ) Sphere formation assay of NFATC2-knockdown and control primary CRC cells. Primary CRC cells were isolated from the cancer tissues of CRC patients (No 1 and 2). The 1st passaged spheres were obtained by suspension culture for 15 days and the number and average diameters of the spheres were counted ( A ). The number of 1st, 2nd, and 3rd passaged spheres was also counted ( B ). ( C ) qRT-PCR analysis of CD44 (top) and CD133 (bottom) in NFATC2-knockdown and control primary CRC cells. Primary CRC cells were isolated from the cancer tissues of CRC patients (No 1 and 2). NFATC2-knockdown and control cells were generated by lentivirus delivery system. ( D ) Western blot analysis of CD44 and CD133 in NFATC2-knockdown and control primary CRC cells. ( E ) Tumorigenesis of NFATC2-knockdown and control primary CRC cells. Data are represented as mean ± SD; *** P <0.001; two-tailed Student’s t -test. Abbreviations: CRC, colorectal cancer; NFATC2, nuclear factor of activated T-cells, cytoplasmic 2; CRC-SCs, colorectal cancer stem cells; qRT-PCR, quantitative real-time polymerase chain reaction.

Article Snippet: CD133 , IB , Mouse monoclonal , , Miltenyi Biotec , 130-092-395 , 100.

Techniques: Knockdown, Tube Formation Assay, Control, Isolation, Suspension, Quantitative RT-PCR, Generated, Western Blot, Two Tailed Test, Real-time Polymerase Chain Reaction

Journal: OncoTargets and therapy

Article Title: NFATC2 is a novel therapeutic target for colorectal cancer stem cells

doi: 10.2147/OTT.S169129

Figure Lengend Snippet: YAP activity is necessary for NFATC2 for maintaining the stemness in CRC cells. ( A , B ) Sphere formation assay of YAP-knockdown NFATC2-overexpressing primary CRC cells and control cells. Primary CRC cells were isolated from the cancer tissues of CRC patients (No 1 and 2). The 1st passaged spheres were obtained by suspension culture for 15 days and the number of the spheres were counted ( A ). The number of 1st, 2nd, and 3rd passaged spheres was also counted ( B ). ( C ) qRT-PCR analysis of CD44 (top) and CD133 (bottom) in YAP-knockdown NFATC2-overexpressing primary CRC cells and control cells. Primary CRC cells were isolated from the cancer tissues of CRC patients (No 1 and 2). YAP-knockdown NFATC2-overexpressing primary CRC cells and control cells were generated by lentivirus delivery system. ( D ) Western blot analysis of CD44 and CD133 in YAP-knockdown NFATC2-overexpressing primary CRC cells and control cells. ( E ) Tumorigenesis of YAP-knockdown NFATC2-overexpressing primary CRC cells and control cells. Data are represented as mean ± SD; *** P <0.001; two-tailed Student’s t -test. Abbreviations: CRC, colorectal cancer; NFATC2, nuclear factor of activated T-cells, cytoplasmic 2; CRC-SCs, colorectal cancer stem cells; qRT-PCR, quantitative real-time polymerase chain reaction.

Article Snippet: CD133 , IB , Mouse monoclonal , , Miltenyi Biotec , 130-092-395 , 100.

Techniques: Activity Assay, Tube Formation Assay, Knockdown, Control, Isolation, Suspension, Quantitative RT-PCR, Generated, Western Blot, Two Tailed Test, Real-time Polymerase Chain Reaction

Journal: OncoTargets and therapy

Article Title: NFATC2 is a novel therapeutic target for colorectal cancer stem cells

doi: 10.2147/OTT.S169129

Figure Lengend Snippet: Primer sequences used in this study

Article Snippet: CD133 , IB , Mouse monoclonal , , Miltenyi Biotec , 130-092-395 , 100.

Techniques: Reverse Transcription

Journal: OncoTargets and therapy

Article Title: NFATC2 is a novel therapeutic target for colorectal cancer stem cells

doi: 10.2147/OTT.S169129

Figure Lengend Snippet: Antibodies used in this study

Article Snippet: CD133 , IB , Mouse monoclonal , , Miltenyi Biotec , 130-092-395 , 100.

Techniques:

Journal: International journal of molecular medicine

Article Title: RACK1 affects glioma cell growth and differentiation through the CNTN2-mediated RTK/Ras/MAPK pathway.

doi: 10.3892/ijmm.2015.2421

Figure Lengend Snippet: Figure 7. Relative protein levels of cell differentiation markers glial fibrillary acidic protein (GFAP) and CD133 in cells following transfection with receptor for activated C kinase 1 (RACK1)-pcDNA3.1 and contactin-2 (CNTN2) siRNA. β-actin was used as an internal control. Lane 1, Vec + Scr si; lane 2, Vec + CNTN2 siRNA (si); lane 3, RACK1 + Scr si; lane 4, RACK1 + CNTN2 si. *P<0.05 and #P<0.01 compared with the Vec + Scr si group; &P<0.05 and $P<0.01 compared with the RACK1 + Scr si group. Vec, pcDNA3.1 vector; Scr si, scrambled siRNA.

Article Snippet: After blocking with 5% bovine serum albumin (Amresco LLC, Solon, OH, USA), the membranes were washed with TBST 3 times, and incubated with primary antibodies, namely rabbit polyclonal antibody to RACK1 (1:800; ab62735), rabbit polyclonal antibody to CNTN2 (1:400; ab68994), rabbit polyclonal antibody to epidermal growth factor receptor (EGFR; 1:800; ab2430), rabbit polyclonal antibody to platelet-derived growth factor receptor, α polypeptide (PDGFRα; 1:500; ab65258), and mouse monoclonal antibody to β-actin (1:2,000; ab6276) (all from Abcam, Cambridge, MA, USA), mouse monoclonal antibody to Ras (1:500; sc-166691), rabbit polyclonal antibody to glial fibrillary acidic protein (GFAP) (1:400; sc-9065) (all from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), rabbit polyclonal antibody to CD133 (1:400; EAP0023; Elabscience, Hubei, China), rabbit monoclonal antibody to ERK1/2 (1:400; #4695), rabbit monoclonal antibody to p-ERK1/2 (Thr202/Tyr204; 1:400; #4377) (Cell Signaling Technology, Inc., Beverly, MA, USA) at 4 ̊C overnight, followed by incubation with HRP-conjugated secondary antibodies, namely goat anti-rabbit IgG HRP (1:2,000; ab6721) and rabbit anti-mouse IgG HRP (1:2,000; ab6728) (both from Abcam) at 37 ̊C for 1 h. The immunocomplexes were visualized by enhanced chemiluminescence (Western Blotting Luminol reagent; Santa Cruz Biotechnology, Inc.).

Techniques: Cell Differentiation, Transfection, Control, Plasmid Preparation