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: Exploration of medicine

Article Title: The association between circulating CD34+CD133+ endothelial progenitor cells and reduced risk of Alzheimer’s disease in the Framingham Heart Study

doi: 10.37349/emed.2024.00216

Figure Lengend Snippet: Examination of the relationship between different circulating progenitor or mature endothelial cells and the risk of AD/dementia by using Cox proportional hazards regression models after adjusting for covariates

Article Snippet: Then, PBMCs were isolated via Ficoll (General Electric) gradient centrifugation, counted, incubated with FcR blocking antibodies (Miltenyi Biotec), labeled with anti-human KDR PE antibody (R&D Systems), anti-human CD34 FITC antibody (BD Biosciences), anti-human CD133 APC antibody (BD Biosciences).

Techniques:

Journal: Exploration of medicine

Article Title: The association between circulating CD34+CD133+ endothelial progenitor cells and reduced risk of Alzheimer’s disease in the Framingham Heart Study

doi: 10.37349/emed.2024.00216

Figure Lengend Snippet: Detailed examination of the relationship between CD34+CD133+ (log-transformed) and the risk of AD/dementia risk

Article Snippet: Then, PBMCs were isolated via Ficoll (General Electric) gradient centrifugation, counted, incubated with FcR blocking antibodies (Miltenyi Biotec), labeled with anti-human KDR PE antibody (R&D Systems), anti-human CD34 FITC antibody (BD Biosciences), anti-human CD133 APC antibody (BD Biosciences).

Techniques:

Journal: Exploration of medicine

Article Title: The association between circulating CD34+CD133+ endothelial progenitor cells and reduced risk of Alzheimer’s disease in the Framingham Heart Study

doi: 10.37349/emed.2024.00216

Figure Lengend Snippet: Characteristics of the study sample based on circulating CD34+CD133+ EPC quartiles

Article Snippet: Then, PBMCs were isolated via Ficoll (General Electric) gradient centrifugation, counted, incubated with FcR blocking antibodies (Miltenyi Biotec), labeled with anti-human KDR PE antibody (R&D Systems), anti-human CD34 FITC antibody (BD Biosciences), anti-human CD133 APC antibody (BD Biosciences).

Techniques:

Journal: Exploration of medicine

Article Title: The association between circulating CD34+CD133+ endothelial progenitor cells and reduced risk of Alzheimer’s disease in the Framingham Heart Study

doi: 10.37349/emed.2024.00216

Figure Lengend Snippet: Survival analyses of circulating CD34+CD133+ cells for AD risk. Circulating the proportions (%) of CD34+CD133+ EPCs were divided into four quartiles based on the cell numbers (Q1 = 0.002–0.021; Q2 = 0.021–0.033; Q3 = 0.033–0.050; and Q4 = 0.050–0.609). Kaplan-Meier survival analysis was conducted to evaluate the survival-free time before AD over 14 years of follow-up to examine the relationship between CD34+CD133+ quartiles and AD incidence for: (A) all of the participants; (B) and (C) stratified based on the genotypes, (B) kirre like nephrin family adhesion molecule 3 (KIRREL3) rs4144611 major homozygotes TT vs. GG + TG; (C) KIRREL3 rs580382 major homozygotes CC vs. TT + CT, and (D) exocyst complex component 6B (EXOC6B) rs61619102 major homozygotes CC vs. GG + GC. P values are shown for statistical significance. Q: quartile

Article Snippet: Then, PBMCs were isolated via Ficoll (General Electric) gradient centrifugation, counted, incubated with FcR blocking antibodies (Miltenyi Biotec), labeled with anti-human KDR PE antibody (R&D Systems), anti-human CD34 FITC antibody (BD Biosciences), anti-human CD133 APC antibody (BD Biosciences).

Techniques:

Journal: Exploration of medicine

Article Title: The association between circulating CD34+CD133+ endothelial progenitor cells and reduced risk of Alzheimer’s disease in the Framingham Heart Study

doi: 10.37349/emed.2024.00216

Figure Lengend Snippet: The relationship between circulating CD34+CD133+ cells and reduced AD risk in the presence of vascular diseases. The FHS participants were stratified into divided based on the CD34+CD133+ EPC proportion quartiles and further divided into those individuals with the absence or presence of different vascular diseases. In the absence of any peripheral and central vascular diseases (A, B), in the presence of CHD (C, D), HTN (E, F), stroke (G, H), silent infarcts (I, J), CMBs (K, L), and high WMHI (M, N), the numbers and percentages of incident AD (A, C, E, G, I, K, and M) and all-cause dementia (B, D, F, H, J, L, and N) across four CD34+CD133+ quartiles were examined by using Chi-squared test with indicated P values. Q: quartile

Article Snippet: Then, PBMCs were isolated via Ficoll (General Electric) gradient centrifugation, counted, incubated with FcR blocking antibodies (Miltenyi Biotec), labeled with anti-human KDR PE antibody (R&D Systems), anti-human CD34 FITC antibody (BD Biosciences), anti-human CD133 APC antibody (BD Biosciences).

Techniques:

Journal: Exploration of medicine

Article Title: The association between circulating CD34+CD133+ endothelial progenitor cells and reduced risk of Alzheimer’s disease in the Framingham Heart Study

doi: 10.37349/emed.2024.00216

Figure Lengend Snippet: Comparisons of brain volumes across circulating CD34+CD133+ EPC quartiles

Article Snippet: Then, PBMCs were isolated via Ficoll (General Electric) gradient centrifugation, counted, incubated with FcR blocking antibodies (Miltenyi Biotec), labeled with anti-human KDR PE antibody (R&D Systems), anti-human CD34 FITC antibody (BD Biosciences), anti-human CD133 APC antibody (BD Biosciences).

Techniques:

Journal: Exploration of medicine

Article Title: The association between circulating CD34+CD133+ endothelial progenitor cells and reduced risk of Alzheimer’s disease in the Framingham Heart Study

doi: 10.37349/emed.2024.00216

Figure Lengend Snippet: GWAS on the interactive effects of genetic factors and circulating CD34+CD133+ cells on AD risk. The FSH analytical pipeline was used to perform GWAS. The interaction between genome-wide SNP dosage and log-transformed proportions (%) of circulating CD34+CD133+ EPCs for AD risk was tested by using the GEEPACK model (logistic regression utilizing generalized estimating equations) adjusting for age, sex, years of education, and the first 10 PCs. As shown in the Manhattan plot, 8 SNPs showed significant interactions with CD34+CD133+ frequency on AD risk (A). Seven of the identified SNPs with a P value < 5.0 × 10 −8 originated from gene KIRREL3 on chromosome 11 (rs580382 and rs4144611 were selected for further analysis), and 1 of them originated from EXOC6B (chromosome 2, rs61619102). Only r 2 ≥ 0.1 and SNPs with MAF ≥ 10% were considered. Manhattan plot (A) was used for visualization to identify the genotypes of two genes ( KIRREL3 and EXOC6B ) that had interactive effects with CD34+CD133+ EPCs for AD risk. Gene structures, surrounding regions, and methylation sites of KIRREL3 (B) and EXOC6B (C) are presented by using LocusZoom

Article Snippet: Then, PBMCs were isolated via Ficoll (General Electric) gradient centrifugation, counted, incubated with FcR blocking antibodies (Miltenyi Biotec), labeled with anti-human KDR PE antibody (R&D Systems), anti-human CD34 FITC antibody (BD Biosciences), anti-human CD133 APC antibody (BD Biosciences).

Techniques: Genome Wide, Transformation Assay, Methylation

Journal: Exploration of medicine

Article Title: The association between circulating CD34+CD133+ endothelial progenitor cells and reduced risk of Alzheimer’s disease in the Framingham Heart Study

doi: 10.37349/emed.2024.00216

Figure Lengend Snippet: Interactions between SNPs of KIRREL3 or EXOC6B and CD34+CD133+ on AD risk in FHS

Article Snippet: Then, PBMCs were isolated via Ficoll (General Electric) gradient centrifugation, counted, incubated with FcR blocking antibodies (Miltenyi Biotec), labeled with anti-human KDR PE antibody (R&D Systems), anti-human CD34 FITC antibody (BD Biosciences), anti-human CD133 APC antibody (BD Biosciences).

Techniques:

Journal: Exploration of medicine

Article Title: The association between circulating CD34+CD133+ endothelial progenitor cells and reduced risk of Alzheimer’s disease in the Framingham Heart Study

doi: 10.37349/emed.2024.00216

Figure Lengend Snippet: The association between circulating CD34+CD133+ cells and the risk of AD in the context of genetic background

Article Snippet: Then, PBMCs were isolated via Ficoll (General Electric) gradient centrifugation, counted, incubated with FcR blocking antibodies (Miltenyi Biotec), labeled with anti-human KDR PE antibody (R&D Systems), anti-human CD34 FITC antibody (BD Biosciences), anti-human CD133 APC antibody (BD Biosciences).

Techniques:

Journal: Respiratory Research

Article Title: Chronic intermittent hypoxia promoted lung cancer stem cell-like properties via enhancing Bach1 expression

doi: 10.1186/s12931-021-01655-6

Figure Lengend Snippet: Primers used for real-time PCR

Article Snippet: A549 and SPCA1 were seeded in 12-well plates at a proper concentration and cultured under Nor or CIH condition for 48 h. At the end of CIH cycles, cells were harvested, filtration and centrifugation, and FITC-labeled anti-CD44 (555478) and APC-labeled anti-CD133 (53276) (Cell Signaling Technologies, USA) were used for surface staining.

Techniques:

Journal: Respiratory Research

Article Title: Chronic intermittent hypoxia promoted lung cancer stem cell-like properties via enhancing Bach1 expression

doi: 10.1186/s12931-021-01655-6

Figure Lengend Snippet: CIH promoted CSC-like properties in NSCLC cells. a Microscopic observation of NSCLC cells. A549 and SPCA1 spheroids were obtained and then cultured under Nor or CIH conditions for 24 h. b qRCP analyses in triplicate of CD44, Sox2, Nanog, Oct4, and CD133 in NSCLC cells. Adherent A549 and SPCA1 were treated with 24 h-Nor or CIH exposure. GADPH expression served as an internal control. c Flow cytometry analyses of the percentage of CD44 + CD133 + cells in A549 or SPCA1 populations. Error bars represent the mean ± SEM of at least triplicate experiments. * P < 0.05, ** P < 0.01. CIH chronic intermittent hypoxia, CSC cancer stem cell

Article Snippet: A549 and SPCA1 were seeded in 12-well plates at a proper concentration and cultured under Nor or CIH condition for 48 h. At the end of CIH cycles, cells were harvested, filtration and centrifugation, and FITC-labeled anti-CD44 (555478) and APC-labeled anti-CD133 (53276) (Cell Signaling Technologies, USA) were used for surface staining.

Techniques: Cell Culture, Expressing, Flow Cytometry

Journal: Respiratory Research

Article Title: Chronic intermittent hypoxia promoted lung cancer stem cell-like properties via enhancing Bach1 expression

doi: 10.1186/s12931-021-01655-6

Figure Lengend Snippet: Knockdown of Bach1 decreased the stemness and mitochondrial ROS accumulation in CIH-treated NSCLCs. Bach1 shRNA or parental negative control (NC) was transfected into A549 or SPCA1. Then the cells were cultured under CIH conditions for 48hrs. Flow cytometry analyses of CD44 + CD133 + cells in Nor or CIH-treated A549 ( a ) or SPCA1 ( b ). c The percentage of CD44 + CD133 + cells in Nor or CIH-treated cells was measured and illustrated. Fluorescence microscopy analysis for localization of mtROS production in A549 ( d ) and SPCA1 ( e ). Nuclei was stained with Hoechst (blue), mitochondria ROS were stained with MitoSOX-red. The merged panels showed the MitoSOX-red positive cells in total cells. f The percentage of MitoSOX positive cells was measured and illustrated. All experiments were performed in triplicate, and data are presented as mean ± SEM. * P < 0.05, ** P < 0.01 compared with the control group

Article Snippet: A549 and SPCA1 were seeded in 12-well plates at a proper concentration and cultured under Nor or CIH condition for 48 h. At the end of CIH cycles, cells were harvested, filtration and centrifugation, and FITC-labeled anti-CD44 (555478) and APC-labeled anti-CD133 (53276) (Cell Signaling Technologies, USA) were used for surface staining.

Techniques: shRNA, Negative Control, Transfection, Cell Culture, Flow Cytometry, Fluorescence, Microscopy, Staining