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recombinant human ccl22 (R&D Systems)

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R&D Systems recombinant human ccl22

Fig. 3. Chemotaxis occurs at much higher rate in KK1 mutant (KK1m) compared to the KK1 revertant (KK1r). (A) Repre sentative timecourse of chemotaxis over 10 h, quantified as % confluence of cells in the top well, normalized to % confluence at t = 0 (left panel). Recombinant human <t>CCL22</t> (500 ng/mL) was added in the bottom well to induce migration. Statistical analysis of the chemotaxis timecourse replicates (n = 3) at the beginning (t = 0 h), middle (t = 5 h), and end (t = 10 h) of the experiment (right panel). Significant differences are calculated and indicated by asterisks as in Fig. 2. (B) Representative images of KK1m and KK1r cell in the top well at t = 10 h.

Recombinant Human Ccl22, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/recombinant+human+ccl22/Recombinant+Human+CCL22%2FMDC+Protein/pm36442790-103-20-23
Average 92 stars, based on 31 article reviews

recombinant human ccl22 - by Bioz Stars, 2026-10

92/100 stars

Images

1) Product Images from "A frequent PLCγ1 mutation in adult T-cell leukemia/lymphoma determines functional properties of the malignant cells."

Article Title: A frequent PLCγ1 mutation in adult T-cell leukemia/lymphoma determines functional properties of the malignant cells.

Journal: Biochimica et biophysica acta. Molecular basis of disease

doi: 10.1016/j.bbadis.2022.166601

Figure Legend Snippet: Fig. 3. Chemotaxis occurs at much higher rate in KK1 mutant (KK1m) compared to the KK1 revertant (KK1r). (A) Repre sentative timecourse of chemotaxis over 10 h, quantified as % confluence of cells in the top well, normalized to % confluence at t = 0 (left panel). Recombinant human CCL22 (500 ng/mL) was added in the bottom well to induce migration. Statistical analysis of the chemotaxis timecourse replicates (n = 3) at the beginning (t = 0 h), middle (t = 5 h), and end (t = 10 h) of the experiment (right panel). Significant differences are calculated and indicated by asterisks as in Fig. 2. (B) Representative images of KK1m and KK1r cell in the top well at t = 10 h.

Techniques Used: Chemotaxis Assay, Mutagenesis, Recombinant, Migration

Fig. 5. Effect of ritonavir and ibrutinib on cell clumping and chemotaxis. (A) Mean rate of clumping (n = 4), in the presence of 100 U/mL IL-2, of KK1m and KK1r lines at the beginning (t = 0 h), middle (t = 20 h), and end (t = 40 h) of the timecourse, normalized to t = 0, with or without ritonavir (left panel) or ibrutinib (IB) (right panel). (B) As in (A) but measuring the average object area of the clumps with and without ritonavir (RITO) (left panel) or ibrutinib (IB) (right panel). (C) Mean rate of chemotaxis (n = 3), induced by 500 ng/mL CCL22, of KK1m cells with and without ritonavir (RITO) (left panel) or ibrutinib (IB) (right panel) at the beginning (t = 0 h), middle (t = 5 h), and end (t = 10 h) of the timecourse, normalized to t = 0. Significant differences are calculated and indicated by asterisks as in Fig. 2.

Figure Legend Snippet: Fig. 5. Effect of ritonavir and ibrutinib on cell clumping and chemotaxis. (A) Mean rate of clumping (n = 4), in the presence of 100 U/mL IL-2, of KK1m and KK1r lines at the beginning (t = 0 h), middle (t = 20 h), and end (t = 40 h) of the timecourse, normalized to t = 0, with or without ritonavir (left panel) or ibrutinib (IB) (right panel). (B) As in (A) but measuring the average object area of the clumps with and without ritonavir (RITO) (left panel) or ibrutinib (IB) (right panel). (C) Mean rate of chemotaxis (n = 3), induced by 500 ng/mL CCL22, of KK1m cells with and without ritonavir (RITO) (left panel) or ibrutinib (IB) (right panel) at the beginning (t = 0 h), middle (t = 5 h), and end (t = 10 h) of the timecourse, normalized to t = 0. Significant differences are calculated and indicated by asterisks as in Fig. 2.

Techniques Used: Chemotaxis Assay

Chemotaxis Assay:

Article Title: A frequent PLCγ1 mutation in adult T-cell leukemia/lymphoma determines functional properties of the malignant cells.
Article Snippet: .. For chemotaxis experiments, growth medium was used without serum to minimize proliferation, with or without the addition of 500 ng/mL recombinant human CCL22 (R&D systems 336-MD-025) in the bottom well to induce migration. ..

Recombinant:

Article Title: Fucoidan inhibits CCL22 production through NF-κB pathway in M2 macrophages: a potential therapeutic strategy for cancer
Article Snippet: .. Recombinant human CCL22, M-CSF, IFN-γ, IL-4 and IL-13 were purchased from R&D Systems (Minneapolis, MN, USA). .. Fucoidan (from Fucus vesiculosus ), PMA, LPS, Wortmannin, SB203580 and BAY 11-7082 were purchased from Sigma-Aldrich (St Louis, MO, USA).

Migration:

Image Search Results

Characterization of double emulsion CCL22 microparticles. ( A ) Representative SEM image demonstrating the porous, spherical particles. ( B ) Size distribution as determined by volume impedance measurements (mean diameter 17.4 ± 7.0 μm). ( C ) Release kinetics of CCL22 over a 21-day period, as determined via in vitro release assay and ELISA. Data are mean ± SEM ( n = 3). ( D ) Biological activity of released CCL22 compared to fresh and denatured CCL22 measured using a cell-based assay. Higher response ratio corresponds to greater bioactivity. Results are mean ± SD ( n = 3).

Journal: Pharmaceutics

Article Title: Encapsulation of Therapeutic, Low-Molecular-Weight Chemokines Using a Single Emulsion, Microfluidic, Continuous Manufacturing Process

doi: 10.3390/pharmaceutics17081056

Figure Lengend Snippet: Characterization of double emulsion CCL22 microparticles. ( A ) Representative SEM image demonstrating the porous, spherical particles. ( B ) Size distribution as determined by volume impedance measurements (mean diameter 17.4 ± 7.0 μm). ( C ) Release kinetics of CCL22 over a 21-day period, as determined via in vitro release assay and ELISA. Data are mean ± SEM ( n = 3). ( D ) Biological activity of released CCL22 compared to fresh and denatured CCL22 measured using a cell-based assay. Higher response ratio corresponds to greater bioactivity. Results are mean ± SD ( n = 3).

Article Snippet: Recombinant human CCL22 (rhCCL22) and recombinant human CCL2 (rhCCL2; Peprotech, Cranberry, NJ, USA) were purchased from Fisher Scientific.

Techniques: Double Emulsion, In Vitro, Release Assay, Enzyme-linked Immunosorbent Assay, Activity Assay, Cell Based Assay

Characterization of single emulsion CCL22 microparticles. ( A ) Representative SEM image demonstrating smooth spherical MPs. ( B ) Size distribution as determined by volume impedance measurements (mean diameter: 20.2 ± 6.0 μm). ( C ) Release kinetics of CCL22 over a 21-day period, as determined via in vitro release assay and ELISA. Data are mean ± SEM ( n = 3). ( D ) Biological activity of released CCL22 compared to fresh and denatured CCL22 through use of a cell-based assay. Results are mean ± SD ( n = 3).

Journal: Pharmaceutics

Article Title: Encapsulation of Therapeutic, Low-Molecular-Weight Chemokines Using a Single Emulsion, Microfluidic, Continuous Manufacturing Process

doi: 10.3390/pharmaceutics17081056

Figure Lengend Snippet: Characterization of single emulsion CCL22 microparticles. ( A ) Representative SEM image demonstrating smooth spherical MPs. ( B ) Size distribution as determined by volume impedance measurements (mean diameter: 20.2 ± 6.0 μm). ( C ) Release kinetics of CCL22 over a 21-day period, as determined via in vitro release assay and ELISA. Data are mean ± SEM ( n = 3). ( D ) Biological activity of released CCL22 compared to fresh and denatured CCL22 through use of a cell-based assay. Results are mean ± SD ( n = 3).

Article Snippet: Recombinant human CCL22 (rhCCL22) and recombinant human CCL2 (rhCCL2; Peprotech, Cranberry, NJ, USA) were purchased from Fisher Scientific.

Techniques: Emulsion, In Vitro, Release Assay, Enzyme-linked Immunosorbent Assay, Activity Assay, Cell Based Assay

Altering initial burst release through changing polymer endcap. ( A ) Representative SEM image of CCL22-MPs manufactured via SE method with hydroxyl-terminated PLGA, demonstrating smooth spherical particles. ( B ) Size distribution as determined via volume impedance measurements (mean diameter: 22.5 ± 7.3 μm). ( C ) Release kinetics of CCL22 over a 21-day period as determined by in vitro release assay and ELISA. Data are mean ± SEM ( n = 3). ( D ) Comparison of initial burst release between CCL22-MPs manufactured with PLGA-COOH vs. PLGA-OH. Results are mean ± SD ( n = 3). A significant difference was identified by a two-tailed independent t -test. * p < 0.05.

Journal: Pharmaceutics

Article Title: Encapsulation of Therapeutic, Low-Molecular-Weight Chemokines Using a Single Emulsion, Microfluidic, Continuous Manufacturing Process

doi: 10.3390/pharmaceutics17081056

Figure Lengend Snippet: Altering initial burst release through changing polymer endcap. ( A ) Representative SEM image of CCL22-MPs manufactured via SE method with hydroxyl-terminated PLGA, demonstrating smooth spherical particles. ( B ) Size distribution as determined via volume impedance measurements (mean diameter: 22.5 ± 7.3 μm). ( C ) Release kinetics of CCL22 over a 21-day period as determined by in vitro release assay and ELISA. Data are mean ± SEM ( n = 3). ( D ) Comparison of initial burst release between CCL22-MPs manufactured with PLGA-COOH vs. PLGA-OH. Results are mean ± SD ( n = 3). A significant difference was identified by a two-tailed independent t -test. * p < 0.05.

Article Snippet: Recombinant human CCL22 (rhCCL22) and recombinant human CCL2 (rhCCL2; Peprotech, Cranberry, NJ, USA) were purchased from Fisher Scientific.

Techniques: Polymer, In Vitro, Release Assay, Enzyme-linked Immunosorbent Assay, Comparison, Two Tailed Test

Microfluidic manufacturing of CCL22 microparticles. ( A ) Representative SEM image demonstrating monodispersed particles with smooth morphology. ( B ) Size distribution as determined via volume impedance measurements (mean diameter: 23.3 ± 4.1 μm). ( C ) Release kinetics of CCL22 over a 21-day period as determined by in vitro release assay and ELISA. Data are mean ± SEM ( n = 3). ( D ) Burst release comparison between CCL22-MPs manufactured via microfluidic (continuous) and homogenization (batch) methods. ( E ) Comparison of protein loading between CCL22-MPs manufactured via microfluidic (continuous) and homogenization (batch) methods. Release and loading characterization results in C-E are presented as mean ± SD ( n = 3). Significant differences in D–E were determined by a two-tailed independent t -test. ** p < 0.01, **** p < 0.0001. ( F ) Biological activity of released CCL22 compared to fresh (unencapsulated) and heat-denatured CCL22, measured via a cell-based assay. Results are mean ± SD ( n = 3).

Journal: Pharmaceutics

Article Title: Encapsulation of Therapeutic, Low-Molecular-Weight Chemokines Using a Single Emulsion, Microfluidic, Continuous Manufacturing Process

doi: 10.3390/pharmaceutics17081056

Figure Lengend Snippet: Microfluidic manufacturing of CCL22 microparticles. ( A ) Representative SEM image demonstrating monodispersed particles with smooth morphology. ( B ) Size distribution as determined via volume impedance measurements (mean diameter: 23.3 ± 4.1 μm). ( C ) Release kinetics of CCL22 over a 21-day period as determined by in vitro release assay and ELISA. Data are mean ± SEM ( n = 3). ( D ) Burst release comparison between CCL22-MPs manufactured via microfluidic (continuous) and homogenization (batch) methods. ( E ) Comparison of protein loading between CCL22-MPs manufactured via microfluidic (continuous) and homogenization (batch) methods. Release and loading characterization results in C-E are presented as mean ± SD ( n = 3). Significant differences in D–E were determined by a two-tailed independent t -test. ** p < 0.01, **** p < 0.0001. ( F ) Biological activity of released CCL22 compared to fresh (unencapsulated) and heat-denatured CCL22, measured via a cell-based assay. Results are mean ± SD ( n = 3).

Article Snippet: Recombinant human CCL22 (rhCCL22) and recombinant human CCL2 (rhCCL2; Peprotech, Cranberry, NJ, USA) were purchased from Fisher Scientific.

Techniques: In Vitro, Release Assay, Enzyme-linked Immunosorbent Assay, Comparison, Homogenization, Two Tailed Test, Activity Assay, Cell Based Assay

rhCCL22-loaded PLGA microparticles exhibit different size distribution based on the manufacturing method. ( a , b ) Volume-weighted size distributions and ( c , d ) representative scanning electron microscopy images of rhCCL22-loaded PLGA microparticles that were prepared by ( a , c ) microfluidics or ( b , d ) conventional methods.

Journal: Pharmaceutics

Article Title: Investigation of the Impact of Manufacturing Methods on Protein-Based Long-Acting Injectable Formulations: A Comparative Assessment for Microfluidics vs. Conventional Methods

doi: 10.3390/pharmaceutics16101264

Figure Lengend Snippet: rhCCL22-loaded PLGA microparticles exhibit different size distribution based on the manufacturing method. ( a , b ) Volume-weighted size distributions and ( c , d ) representative scanning electron microscopy images of rhCCL22-loaded PLGA microparticles that were prepared by ( a , c ) microfluidics or ( b , d ) conventional methods.

Article Snippet: Recombinant human CCL22 (rhCCL22; 69 a.a.) was purchased from PeproTech (Cranbury, NJ, USA).

Techniques: Electron Microscopy

Representative scanning electron microscopy images demonstrate the surface morphology and inner porosity differences for rhCCL22-loaded PLGA microparticles. PLGA microparticles were prepared by ( a , c ) microfluidics and ( b , d ) conventional methods. Arrows in ( d ) highlight some examples of the inner pore connections.

Journal: Pharmaceutics

Article Title: Investigation of the Impact of Manufacturing Methods on Protein-Based Long-Acting Injectable Formulations: A Comparative Assessment for Microfluidics vs. Conventional Methods

doi: 10.3390/pharmaceutics16101264

Figure Lengend Snippet: Representative scanning electron microscopy images demonstrate the surface morphology and inner porosity differences for rhCCL22-loaded PLGA microparticles. PLGA microparticles were prepared by ( a , c ) microfluidics and ( b , d ) conventional methods. Arrows in ( d ) highlight some examples of the inner pore connections.

Article Snippet: Recombinant human CCL22 (rhCCL22; 69 a.a.) was purchased from PeproTech (Cranbury, NJ, USA).

Techniques: Electron Microscopy

rhCCL22 encapsulation (%) in the PLGA microparticles shows a significant difference depending on the manufacturing process: microfluidics vs. conventional methods. ** p ≤ 0.01.

Journal: Pharmaceutics

Article Title: Investigation of the Impact of Manufacturing Methods on Protein-Based Long-Acting Injectable Formulations: A Comparative Assessment for Microfluidics vs. Conventional Methods

doi: 10.3390/pharmaceutics16101264

Figure Lengend Snippet: rhCCL22 encapsulation (%) in the PLGA microparticles shows a significant difference depending on the manufacturing process: microfluidics vs. conventional methods. ** p ≤ 0.01.

Article Snippet: Recombinant human CCL22 (rhCCL22; 69 a.a.) was purchased from PeproTech (Cranbury, NJ, USA).

Techniques: Encapsulation

Journal: Pharmaceutics

Article Title: Investigation of the Impact of Manufacturing Methods on Protein-Based Long-Acting Injectable Formulations: A Comparative Assessment for Microfluidics vs. Conventional Methods

doi: 10.3390/pharmaceutics16101264

Figure Lengend Snippet: rhCCL22 release kinetics from PLGA microparticles demonstrates different release kinetics according to the manufacturing method. ( a ) Microparticles prepared by the microfluidics method showed a zero-order delayed (~2 days) release profile. ( b ) Microparticles prepared by the conventional method presented a zero-order release profile.

Article Snippet: Recombinant human CCL22 (rhCCL22; 69 a.a.) was purchased from PeproTech (Cranbury, NJ, USA).

Techniques:

Batch-to-batch variations in the size distribution of rhCCL22-loaded PLGA microparticles when PLGA microparticles were prepared by ( a ) microfluidics and ( b ) conventional methods.

Journal: Pharmaceutics

Article Title: Investigation of the Impact of Manufacturing Methods on Protein-Based Long-Acting Injectable Formulations: A Comparative Assessment for Microfluidics vs. Conventional Methods

doi: 10.3390/pharmaceutics16101264

Figure Lengend Snippet: Batch-to-batch variations in the size distribution of rhCCL22-loaded PLGA microparticles when PLGA microparticles were prepared by ( a ) microfluidics and ( b ) conventional methods.

Article Snippet: Recombinant human CCL22 (rhCCL22; 69 a.a.) was purchased from PeproTech (Cranbury, NJ, USA).

Techniques:

Batch-to-batch variations in terms of rhCCL22 encapsulation (%) in PLGA microparticles regarding the manufacturing method. rhCCL22-loaded PLGA microparticles were prepared by ( a ) microfluidics and ( b ) conventional methods. * p ≤ 0.05, *** p ≤ 0.001, ns indicates non-significant difference.

Journal: Pharmaceutics

Article Title: Investigation of the Impact of Manufacturing Methods on Protein-Based Long-Acting Injectable Formulations: A Comparative Assessment for Microfluidics vs. Conventional Methods

doi: 10.3390/pharmaceutics16101264

Figure Lengend Snippet: Batch-to-batch variations in terms of rhCCL22 encapsulation (%) in PLGA microparticles regarding the manufacturing method. rhCCL22-loaded PLGA microparticles were prepared by ( a ) microfluidics and ( b ) conventional methods. * p ≤ 0.05, *** p ≤ 0.001, ns indicates non-significant difference.

Article Snippet: Recombinant human CCL22 (rhCCL22; 69 a.a.) was purchased from PeproTech (Cranbury, NJ, USA).

Techniques: Encapsulation

Batch-to-batch variations in terms of rhCCL22 release kinetics from PLGA microparticles. rhCCL22-loaded PLGA microparticles were prepared by ( a ) microfluidics and ( b ) conventional methods.

Journal: Pharmaceutics

Article Title: Investigation of the Impact of Manufacturing Methods on Protein-Based Long-Acting Injectable Formulations: A Comparative Assessment for Microfluidics vs. Conventional Methods

doi: 10.3390/pharmaceutics16101264

Figure Lengend Snippet: Batch-to-batch variations in terms of rhCCL22 release kinetics from PLGA microparticles. rhCCL22-loaded PLGA microparticles were prepared by ( a ) microfluidics and ( b ) conventional methods.

Article Snippet: Recombinant human CCL22 (rhCCL22; 69 a.a.) was purchased from PeproTech (Cranbury, NJ, USA).

Techniques:

Journal: Journal of Cancer

Article Title: Activation of the CCL22/CCR4 causing EMT process remodeling under EZH2-mediated epigenetic regulation in cervical carcinoma

doi: 10.7150/jca.101881

Figure Lengend Snippet: Primer sequences used in the study

Article Snippet: Recombinant human CCL22 (NBP2-34977, NOVUS, USA) was reconstituted according to instructions provided by the manufacturer.

Techniques: Sequencing, Control

Hypomethylation states of CCL22 and CCR4 caused overexpression of CCL22 and CCR4 in CC . ( A ) An overview of mRNA levels of CCL22 and CCR4 in CC based on GEPIA database. ( B ) The mRNA levels of CCL22 and CCR4 in CC (Ca) (n=32) and normal cervical tissues (NC) (n=32) detected by RT-qPCR. ( C ) The correction between CCL22 and CCR4 in CC, analyzed by GEPIA database. ( D ) Predicted CpG islands in the promoter regions of CCL22 and CCR4 . Numbers indicate the positions in bp relative to the transcription start site. The blue region represents the CpG islands and the red vertical bars are the CpG loci in these input sequences. ( E and F ) DNA Methylation level of CCL22 and CCR4 promoter regions in CC (Ca) (n=9) and NC (n=9) detected by MS-PCR. MS-PCR images of 4 representative samples are shown from each group. ( G ) Detection of CCL22 and CCR4 promoter DNA methylation status by MS-PCR in SiHa, Hela and C33A cells; (M: methylated, U: unmethylated). ( H ) Relative mRNA expression of CCL22 and CCR4 in SiHa, HeLa and C33A cells after treatment with different concentrations of 5-Aza-CdR.

Journal: Journal of Cancer

Article Title: Activation of the CCL22/CCR4 causing EMT process remodeling under EZH2-mediated epigenetic regulation in cervical carcinoma

doi: 10.7150/jca.101881

Figure Lengend Snippet: Hypomethylation states of CCL22 and CCR4 caused overexpression of CCL22 and CCR4 in CC . ( A ) An overview of mRNA levels of CCL22 and CCR4 in CC based on GEPIA database. ( B ) The mRNA levels of CCL22 and CCR4 in CC (Ca) (n=32) and normal cervical tissues (NC) (n=32) detected by RT-qPCR. ( C ) The correction between CCL22 and CCR4 in CC, analyzed by GEPIA database. ( D ) Predicted CpG islands in the promoter regions of CCL22 and CCR4 . Numbers indicate the positions in bp relative to the transcription start site. The blue region represents the CpG islands and the red vertical bars are the CpG loci in these input sequences. ( E and F ) DNA Methylation level of CCL22 and CCR4 promoter regions in CC (Ca) (n=9) and NC (n=9) detected by MS-PCR. MS-PCR images of 4 representative samples are shown from each group. ( G ) Detection of CCL22 and CCR4 promoter DNA methylation status by MS-PCR in SiHa, Hela and C33A cells; (M: methylated, U: unmethylated). ( H ) Relative mRNA expression of CCL22 and CCR4 in SiHa, HeLa and C33A cells after treatment with different concentrations of 5-Aza-CdR.

Article Snippet: Recombinant human CCL22 (NBP2-34977, NOVUS, USA) was reconstituted according to instructions provided by the manufacturer.

Techniques: Over Expression, Quantitative RT-PCR, DNA Methylation Assay, Methylation, Expressing

DNMT3A reactivated the CCL22 and CCR4 expression through decreasing promoters' DNA methylation . ( A ) Detection of the expression of DNMT3A , CCL22 and CCR4 in DNMT3A specific siRNA transfected SiHa and HeLa cells by RT-qPCR. ( B ) Detection of the DNA methylation level of CCL22 and CCR4 in DNMT3A specific siRNA transfected SiHa and HeLa cells by MS-qPCR. ( C ) Schematic representation of the 4 regions of the CCL22 and CCR4 promoter regions amplified in the chromatin immunoprecipitation (ChIP)‑quantitative PCR (qPCR) experiment. ( D and E ) Chromatin was cross-linked, fragmented and immunoprecipitated with either IgG (mock) or anti-DNMT3A ChIP-grade antibody and the purified DNA was used to amplify with respective primer pairs for indicated four regions in the CCL22 and CCR4 promoter regions in qPCR. The enrichment of DNMT3A on CCL22 and CCR4 promoter regions relative to IgG in SiHa and HeLa cells, and H3 against RPL30 was used as positive control.

Journal: Journal of Cancer

Article Title: Activation of the CCL22/CCR4 causing EMT process remodeling under EZH2-mediated epigenetic regulation in cervical carcinoma

doi: 10.7150/jca.101881

Figure Lengend Snippet: DNMT3A reactivated the CCL22 and CCR4 expression through decreasing promoters' DNA methylation . ( A ) Detection of the expression of DNMT3A , CCL22 and CCR4 in DNMT3A specific siRNA transfected SiHa and HeLa cells by RT-qPCR. ( B ) Detection of the DNA methylation level of CCL22 and CCR4 in DNMT3A specific siRNA transfected SiHa and HeLa cells by MS-qPCR. ( C ) Schematic representation of the 4 regions of the CCL22 and CCR4 promoter regions amplified in the chromatin immunoprecipitation (ChIP)‑quantitative PCR (qPCR) experiment. ( D and E ) Chromatin was cross-linked, fragmented and immunoprecipitated with either IgG (mock) or anti-DNMT3A ChIP-grade antibody and the purified DNA was used to amplify with respective primer pairs for indicated four regions in the CCL22 and CCR4 promoter regions in qPCR. The enrichment of DNMT3A on CCL22 and CCR4 promoter regions relative to IgG in SiHa and HeLa cells, and H3 against RPL30 was used as positive control.

Article Snippet: Recombinant human CCL22 (NBP2-34977, NOVUS, USA) was reconstituted according to instructions provided by the manufacturer.

Techniques: Expressing, DNA Methylation Assay, Transfection, Quantitative RT-PCR, Amplification, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Immunoprecipitation, Purification, Positive Control

Inhibition of EZH2 promoted DNMT3A in cervical cancer cells with methylated the promoter regions of CCL22 and CCR4 . ( A ) Detection of the expression of EZH2, H3K27me3 and DNMT3A in EZH2 knocked-down or specific siRNA transfected or DZNep treated SiHa and HeLa cells by western blotting. ( B ) Schematic representation of the 4 regions of the DNMT3A promoter region amplified in the chromatin immunoprecipitation (ChIP)‑quantitative PCR (qPCR) experiment. ( C ) Chromatin was cross-linked, fragmented and immunoprecipitated with either IgG (mock) or anti-EZH2 and H3K27me3 ChIP-grade antibody and the purified DNA was used to amplify with respective primer pairs for indicated four regions in the DNMT3A promoter region in qPCR. The enrichment of EZH2 and H3K27me3 on DNMT3A promoter region relative to IgG in SiHa cell, and H3 against RPL30 was used as positive control. ( D , F and H ) The mRNA expression of EZH2 , DNMT3A and CCL22 - CCR4 in EZH2 knocked-down or specific siRNA transfected or DZNep treated SiHa and HeLa cells by RT-qPCR. ( E , G and I ) Detection of the methylation level of CCL22 and CCR4 in EZH2 knocked-down or specific siRNA transfected or DZNep treated SiHa and HeLa cells by MS-qPCR.

Journal: Journal of Cancer

Article Title: Activation of the CCL22/CCR4 causing EMT process remodeling under EZH2-mediated epigenetic regulation in cervical carcinoma

doi: 10.7150/jca.101881

Figure Lengend Snippet: Inhibition of EZH2 promoted DNMT3A in cervical cancer cells with methylated the promoter regions of CCL22 and CCR4 . ( A ) Detection of the expression of EZH2, H3K27me3 and DNMT3A in EZH2 knocked-down or specific siRNA transfected or DZNep treated SiHa and HeLa cells by western blotting. ( B ) Schematic representation of the 4 regions of the DNMT3A promoter region amplified in the chromatin immunoprecipitation (ChIP)‑quantitative PCR (qPCR) experiment. ( C ) Chromatin was cross-linked, fragmented and immunoprecipitated with either IgG (mock) or anti-EZH2 and H3K27me3 ChIP-grade antibody and the purified DNA was used to amplify with respective primer pairs for indicated four regions in the DNMT3A promoter region in qPCR. The enrichment of EZH2 and H3K27me3 on DNMT3A promoter region relative to IgG in SiHa cell, and H3 against RPL30 was used as positive control. ( D , F and H ) The mRNA expression of EZH2 , DNMT3A and CCL22 - CCR4 in EZH2 knocked-down or specific siRNA transfected or DZNep treated SiHa and HeLa cells by RT-qPCR. ( E , G and I ) Detection of the methylation level of CCL22 and CCR4 in EZH2 knocked-down or specific siRNA transfected or DZNep treated SiHa and HeLa cells by MS-qPCR.

Article Snippet: Recombinant human CCL22 (NBP2-34977, NOVUS, USA) was reconstituted according to instructions provided by the manufacturer.

Techniques: Inhibition, Methylation, Expressing, Transfection, Western Blot, Amplification, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Immunoprecipitation, Purification, Positive Control, Quantitative RT-PCR

Effect of CCL22-CCR4 on migration of CC cells . ( A ) The migratory potential of SiHa and HeLa cells which added recombinant human CCL22 protein or neutralization CCL22 antibody and the respective control cells was analyzed by the transwell cell migration assay. Number of migratory cells was shown as means ± standard error from three independent experiments using triplicate measurements and statistically analyzed with Student's t-test in each experiment. Magnification, ×200. ( B ) The expression of EMT-related proteins in SiHa or HeLa cells which added recombinant human CCL22 protein or neutralization CCL22 antibody and the respective control cells was determined by western blotting and the gray level analysis of the protein levels of EMT-related proteins.

Journal: Journal of Cancer

Article Title: Activation of the CCL22/CCR4 causing EMT process remodeling under EZH2-mediated epigenetic regulation in cervical carcinoma

doi: 10.7150/jca.101881

Figure Lengend Snippet: Effect of CCL22-CCR4 on migration of CC cells . ( A ) The migratory potential of SiHa and HeLa cells which added recombinant human CCL22 protein or neutralization CCL22 antibody and the respective control cells was analyzed by the transwell cell migration assay. Number of migratory cells was shown as means ± standard error from three independent experiments using triplicate measurements and statistically analyzed with Student's t-test in each experiment. Magnification, ×200. ( B ) The expression of EMT-related proteins in SiHa or HeLa cells which added recombinant human CCL22 protein or neutralization CCL22 antibody and the respective control cells was determined by western blotting and the gray level analysis of the protein levels of EMT-related proteins.

Article Snippet: Recombinant human CCL22 (NBP2-34977, NOVUS, USA) was reconstituted according to instructions provided by the manufacturer.

Techniques: Migration, Recombinant, Neutralization, Control, Cell Migration Assay, Expressing, Western Blot

Inhibition EZH2 represses migration in CC cells through downregulating CCL22-CCR4 . ( A ) The migratory potential of EZH2 knocked-down SiHa or HeLa cells and the respective control cells was analyzed by the transwell cell migration assay. Number of migratory cells was shown as means ± standard error from three independent experiments using triplicate measurements and statistically analyzed with Student's t-test in each experiment. Magnification, ×200. ( B ) The migratory potential of DNMT3A specific siRNA transfected SiHa and HeLa cells and the respective control cells was analyzed by the transwell cell migration assay. Number of migratory cells was shown as means ± standard error from three independent experiments using triplicate measurements and statistically analyzed with Student's t-test in each experiment. Magnification, ×200. ( C ) The expression of EMT-related proteins in EZH2 knocked-down or DZNep treated SiHa and HeLa cells were determined by western blotting and the gray level analysis of the protein levels of EMT-related proteins. ( D ) The expression of EMT-related proteins in DNMT3A specific siRNA transfected SiHa and HeLa cells were determined by western blotting and the gray level analysis of the protein levels of EMT-related proteins and DNMT3A.

Journal: Journal of Cancer

Article Title: Activation of the CCL22/CCR4 causing EMT process remodeling under EZH2-mediated epigenetic regulation in cervical carcinoma

doi: 10.7150/jca.101881

Figure Lengend Snippet: Inhibition EZH2 represses migration in CC cells through downregulating CCL22-CCR4 . ( A ) The migratory potential of EZH2 knocked-down SiHa or HeLa cells and the respective control cells was analyzed by the transwell cell migration assay. Number of migratory cells was shown as means ± standard error from three independent experiments using triplicate measurements and statistically analyzed with Student's t-test in each experiment. Magnification, ×200. ( B ) The migratory potential of DNMT3A specific siRNA transfected SiHa and HeLa cells and the respective control cells was analyzed by the transwell cell migration assay. Number of migratory cells was shown as means ± standard error from three independent experiments using triplicate measurements and statistically analyzed with Student's t-test in each experiment. Magnification, ×200. ( C ) The expression of EMT-related proteins in EZH2 knocked-down or DZNep treated SiHa and HeLa cells were determined by western blotting and the gray level analysis of the protein levels of EMT-related proteins. ( D ) The expression of EMT-related proteins in DNMT3A specific siRNA transfected SiHa and HeLa cells were determined by western blotting and the gray level analysis of the protein levels of EMT-related proteins and DNMT3A.

Article Snippet: Recombinant human CCL22 (NBP2-34977, NOVUS, USA) was reconstituted according to instructions provided by the manufacturer.

Techniques: Inhibition, Migration, Control, Cell Migration Assay, Transfection, Expressing, Western Blot

CCL22-CCCR4 promotes migration in CC cells . ( A ) The migratory potential of EZH2 knocked-down SiHa or HeLa cells with or without CCL22 and the respective control cells was analyzed by the transwell cell migration assay. Number of migratory cells was shown as means ± standard error from three independent experiments using triplicate measurements and statistically analyzed with Student's t-test in each experiment. Magnification, ×200. ( B ) The expression of EMT-related proteins in EZH2 knocked-down SiHa and HeLa cells with or without CCL22 were determined by western blotting. ( C ) The migratory potential of DNMT3A specific siRNA transfected SiHa and HeLa cells with or without anti-CCL22 and the respective control cells was analyzed by the transwell cell migration assay. Number of migratory cells was shown as means ± standard error from three independent experiments using triplicate measurements and statistically analyzed with Student's t-test in each experiment. Magnification, ×200. ( D ) The expression of EMT-related proteins in DNMT3A specific siRNA transfected SiHa and HeLa cells with or without anti-CCL22 determined by western blotting.

Journal: Journal of Cancer

Article Title: Activation of the CCL22/CCR4 causing EMT process remodeling under EZH2-mediated epigenetic regulation in cervical carcinoma

doi: 10.7150/jca.101881

Figure Lengend Snippet: CCL22-CCCR4 promotes migration in CC cells . ( A ) The migratory potential of EZH2 knocked-down SiHa or HeLa cells with or without CCL22 and the respective control cells was analyzed by the transwell cell migration assay. Number of migratory cells was shown as means ± standard error from three independent experiments using triplicate measurements and statistically analyzed with Student's t-test in each experiment. Magnification, ×200. ( B ) The expression of EMT-related proteins in EZH2 knocked-down SiHa and HeLa cells with or without CCL22 were determined by western blotting. ( C ) The migratory potential of DNMT3A specific siRNA transfected SiHa and HeLa cells with or without anti-CCL22 and the respective control cells was analyzed by the transwell cell migration assay. Number of migratory cells was shown as means ± standard error from three independent experiments using triplicate measurements and statistically analyzed with Student's t-test in each experiment. Magnification, ×200. ( D ) The expression of EMT-related proteins in DNMT3A specific siRNA transfected SiHa and HeLa cells with or without anti-CCL22 determined by western blotting.

Article Snippet: Recombinant human CCL22 (NBP2-34977, NOVUS, USA) was reconstituted according to instructions provided by the manufacturer.

Techniques: Migration, Control, Cell Migration Assay, Expressing, Western Blot, Transfection

Epigenetic modulation of CCL22-CCR4 mediated by EZH2 in vivo . ( A ) SiHa-shEZH2 and HeLa-shEZH2 tumor xenografts in nude mice. ( B ) The tumors weight formed from SiHa-shEZH2 and HeLa-shEZH2. ( C ) Tumors formed from SiHa-shEZH2 and HeLa-shEZH2 cells as well as tumor growth curves. ( D , F and G ) Western blotting and RT-qPCR results of EZH2, H3K27me3, DNMT3A, CCL22-CCR4 and EMT-related proteins in SiHa-shEZH2 and HeLa-shEZH2 cells formed tumors. ( E ) The methylation level of CCL22 and CCR4 promoter regions were monitored by MS-qPCR in tumor tissues. ( H - K ) Chromatin was cross‑linked, fragmented and immunoprecipitated with either IgG (mock) or anti‑EZH2, H3K27me3 and DNMT3A ChIP‑grade antibody and the purified DNA was used to amplify with respective primer pairs for the indicated 4 regions in the DNMT3A , CCL22 - CCR4 promoters in qPCR. The enrichment of EZH2 and H3K27me3 on DNMT3A , CCL22 - CCR4 promoters and the enrichment of DNMT3A CCL22 - CCR4 promoters on relative to IgG in tumor tissues.

Journal: Journal of Cancer

Article Title: Activation of the CCL22/CCR4 causing EMT process remodeling under EZH2-mediated epigenetic regulation in cervical carcinoma

doi: 10.7150/jca.101881

Figure Lengend Snippet: Epigenetic modulation of CCL22-CCR4 mediated by EZH2 in vivo . ( A ) SiHa-shEZH2 and HeLa-shEZH2 tumor xenografts in nude mice. ( B ) The tumors weight formed from SiHa-shEZH2 and HeLa-shEZH2. ( C ) Tumors formed from SiHa-shEZH2 and HeLa-shEZH2 cells as well as tumor growth curves. ( D , F and G ) Western blotting and RT-qPCR results of EZH2, H3K27me3, DNMT3A, CCL22-CCR4 and EMT-related proteins in SiHa-shEZH2 and HeLa-shEZH2 cells formed tumors. ( E ) The methylation level of CCL22 and CCR4 promoter regions were monitored by MS-qPCR in tumor tissues. ( H - K ) Chromatin was cross‑linked, fragmented and immunoprecipitated with either IgG (mock) or anti‑EZH2, H3K27me3 and DNMT3A ChIP‑grade antibody and the purified DNA was used to amplify with respective primer pairs for the indicated 4 regions in the DNMT3A , CCL22 - CCR4 promoters in qPCR. The enrichment of EZH2 and H3K27me3 on DNMT3A , CCL22 - CCR4 promoters and the enrichment of DNMT3A CCL22 - CCR4 promoters on relative to IgG in tumor tissues.

Article Snippet: Recombinant human CCL22 (NBP2-34977, NOVUS, USA) was reconstituted according to instructions provided by the manufacturer.

Techniques: In Vivo, Western Blot, Quantitative RT-PCR, Methylation, Immunoprecipitation, Purification

The pathway of EZH2 regulated CCL22-CCR4 expression through epigenetic modification causing EMT remodeling.

Journal: Journal of Cancer

Article Title: Activation of the CCL22/CCR4 causing EMT process remodeling under EZH2-mediated epigenetic regulation in cervical carcinoma

doi: 10.7150/jca.101881

Figure Lengend Snippet: The pathway of EZH2 regulated CCL22-CCR4 expression through epigenetic modification causing EMT remodeling.

Article Snippet: Recombinant human CCL22 (NBP2-34977, NOVUS, USA) was reconstituted according to instructions provided by the manufacturer.

Techniques: Expressing, Modification

Journal: Biochimica et biophysica acta. Molecular basis of disease

Article Title: A frequent PLCγ1 mutation in adult T-cell leukemia/lymphoma determines functional properties of the malignant cells.

doi: 10.1016/j.bbadis.2022.166601

Figure Lengend Snippet: Fig. 3. Chemotaxis occurs at much higher rate in KK1 mutant (KK1m) compared to the KK1 revertant (KK1r). (A) Repre sentative timecourse of chemotaxis over 10 h, quantified as % confluence of cells in the top well, normalized to % confluence at t = 0 (left panel). Recombinant human CCL22 (500 ng/mL) was added in the bottom well to induce migration. Statistical analysis of the chemotaxis timecourse replicates (n = 3) at the beginning (t = 0 h), middle (t = 5 h), and end (t = 10 h) of the experiment (right panel). Significant differences are calculated and indicated by asterisks as in Fig. 2. (B) Representative images of KK1m and KK1r cell in the top well at t = 10 h.

Article Snippet: For chemotaxis experiments, growth medium was used without serum to minimize proliferation, with or without the addition of 500 ng/mL recombinant human CCL22 (R&D systems 336-MD-025) in the bottom well to induce migration.

Techniques: Chemotaxis Assay, Mutagenesis, Recombinant, Migration

Journal: Biochimica et biophysica acta. Molecular basis of disease

Article Title: A frequent PLCγ1 mutation in adult T-cell leukemia/lymphoma determines functional properties of the malignant cells.

doi: 10.1016/j.bbadis.2022.166601

Figure Lengend Snippet: Fig. 5. Effect of ritonavir and ibrutinib on cell clumping and chemotaxis. (A) Mean rate of clumping (n = 4), in the presence of 100 U/mL IL-2, of KK1m and KK1r lines at the beginning (t = 0 h), middle (t = 20 h), and end (t = 40 h) of the timecourse, normalized to t = 0, with or without ritonavir (left panel) or ibrutinib (IB) (right panel). (B) As in (A) but measuring the average object area of the clumps with and without ritonavir (RITO) (left panel) or ibrutinib (IB) (right panel). (C) Mean rate of chemotaxis (n = 3), induced by 500 ng/mL CCL22, of KK1m cells with and without ritonavir (RITO) (left panel) or ibrutinib (IB) (right panel) at the beginning (t = 0 h), middle (t = 5 h), and end (t = 10 h) of the timecourse, normalized to t = 0. Significant differences are calculated and indicated by asterisks as in Fig. 2.

Techniques: Chemotaxis Assay

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recombinant human ccl22 (R&D Systems)

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