Journal: Oncotarget

Article Title: CCR4 promotes metastasis via ERK/NF-κB/MMP13 pathway and acts downstream of TNF-α in colorectal cancer

doi: 10.18632/oncotarget.10256

Figure Lengend Snippet: ( A ) qRT-PCR analysis showing CCR4 expression in 16 paired CRC samples that were randomly selected from the 116 CRC cases, by random numbers generated with SAS software. ( B ) Representative western blot images of CCR4 expression in 16 paired CRC samples. ( C ) Quantification of relative grey value of bands compared with GAPDH, as detected by western blot. ( D ) CCR4 expression level in tumor tissues and the paired normal tissues was evaluated by immunohistochemical staining with tissue microarray. ( E ) CRC patients with positive expression of CCR4 presented with worse overall survival, and disease free survival compared with that of negative expression of CCR4. Data represent the mean ± SD and are representative of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: pGLV-GFP-CCR4 lentiviral vector and four shRNA plasmids targeting different regions of CCR4 mRNA were purchased from Genepharma (Shanghai, China).

Techniques: Quantitative RT-PCR, Expressing, Generated, Software, Western Blot, Immunohistochemical staining, Staining, Microarray

Journal: Oncotarget

Article Title: CCR4 promotes metastasis via ERK/NF-κB/MMP13 pathway and acts downstream of TNF-α in colorectal cancer

doi: 10.18632/oncotarget.10256

Figure Lengend Snippet: Relationship between CCR4 expression level and clinicopathologic variables in 116 CRC patients

Article Snippet: pGLV-GFP-CCR4 lentiviral vector and four shRNA plasmids targeting different regions of CCR4 mRNA were purchased from Genepharma (Shanghai, China).

Techniques: Expressing

Journal: Oncotarget

Article Title: CCR4 promotes metastasis via ERK/NF-κB/MMP13 pathway and acts downstream of TNF-α in colorectal cancer

doi: 10.18632/oncotarget.10256

Figure Lengend Snippet: ( A ) CCR4 expression in eight CRC cell lines detected by western blot. ( B ) SW1116 and SW480 cells transfected with pcDNA-CCR4 and sh-CCR4, respectively, were subject to western blot. ( C ) Invasive behavior was evaluated using matrigel invasion assays after overexpression or knockdown of CCR4 in SW1116 or SW480 (magnification, ×200). ( D ) The migratory capacity of SW1116/CCR4 and SW480/sh-CCR4 cells was analyzed by wound-healing assay.

Article Snippet: pGLV-GFP-CCR4 lentiviral vector and four shRNA plasmids targeting different regions of CCR4 mRNA were purchased from Genepharma (Shanghai, China).

Techniques: Expressing, Western Blot, Transfection, Over Expression, Wound Healing Assay

Journal: Oncotarget

Article Title: CCR4 promotes metastasis via ERK/NF-κB/MMP13 pathway and acts downstream of TNF-α in colorectal cancer

doi: 10.18632/oncotarget.10256

Figure Lengend Snippet: ( A ) HT29 and SW620 cells transfected with pcDNA-CCR4 and sh-CCR4, respectively, were tested by western blot. ( B ) Matrigel invasion assays were performed to assess invasive ability of HT29/CCR4 and SW620/sh-CCR4 (magnification, ×200). ( C ) Wound-healing assay shows a significant increase or decrease in healing rate of the scramble wound in HT29/CCR4 and SW620/sh-CCR4, respectively. ( D ) Representative images of hematoxylin & eosin staining of liver tissue sections. Black arrows indicated liver metastasis. ( E ) Number of metastasis in the liver. ( F ) Number of tumors on the surface of liver given orthotopic implantation of xenograft. Data represent the mean ± SD and are representative of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: pGLV-GFP-CCR4 lentiviral vector and four shRNA plasmids targeting different regions of CCR4 mRNA were purchased from Genepharma (Shanghai, China).

Techniques: Transfection, Western Blot, Wound Healing Assay, Staining

Journal: Oncotarget

Article Title: CCR4 promotes metastasis via ERK/NF-κB/MMP13 pathway and acts downstream of TNF-α in colorectal cancer

doi: 10.18632/oncotarget.10256

Figure Lengend Snippet: ( A ) Five metastasis-related genes (VEGF, MMP13, CDH1, IL1B, ITGA7) showed a more than 2-fold mRNA differential expression in PCR array. ( B ) The effect of MMP13 knockdown in SW1116/CCR4 and SW480 cells, detected by western blot. Densitometry represents the expression of the proteins relative to GAPDH. ( C ) Results of invasion assays showed the inhibitory roles of si-MMP13 or CL82198 on SW1116/CCR4 and SW480 cells (magnification, ×200). ( D ) Representative images of MMP13 staining in the cohort of 116 CRC tissues. ( E ) Expression correlation of CCR4 and MMP13 was analyzed in 116 CRC patients using IHC.

Article Snippet: pGLV-GFP-CCR4 lentiviral vector and four shRNA plasmids targeting different regions of CCR4 mRNA were purchased from Genepharma (Shanghai, China).

Techniques: Expressing, Western Blot, Staining

Journal: Oncotarget

Article Title: CCR4 promotes metastasis via ERK/NF-κB/MMP13 pathway and acts downstream of TNF-α in colorectal cancer

doi: 10.18632/oncotarget.10256

Figure Lengend Snippet: ( A ) p-ERK, p-p38, p-JNK, p-Akt, p-p65 and MMP13 expressions were determined by western blot analysis. Densitometry represents the expression of the proteins relative to GAPDH. ( B ) p-p65, β-catenin, Runx2, and MMP13 expressions were analyzed using western blot. Densitometry represents the expression of the proteins relative to GAPDH. ( C ) Results of invasion assays showed the inhibitory roles of U0126 or TCPA-1 on SW1116/CCR4 and SW480 cells (magnification, ×200). ( D ) Representative images of IHC staining of p-ERK, p-p65 and MMP13 in the CRC tissues of orthotopic implantation model in nude mice. Scare bars = 50 μm. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: pGLV-GFP-CCR4 lentiviral vector and four shRNA plasmids targeting different regions of CCR4 mRNA were purchased from Genepharma (Shanghai, China).

Techniques: Western Blot, Expressing, Immunohistochemistry

Journal: Oncotarget

Article Title: CCR4 promotes metastasis via ERK/NF-κB/MMP13 pathway and acts downstream of TNF-α in colorectal cancer

doi: 10.18632/oncotarget.10256

Figure Lengend Snippet: ( A ) Cells were incubated with different doses of TNF-α for 12 h, and p-p65, CCR4 or MMP13 expression is examined by western blot. ( B ) Representative images of IHC staining of TNF-α in tissue microarray. ( C ) Prognostic values of CCR4 combined with TNF-α. ( D ) DNA fragments pulled down with NF-κB antibody from SW1116 cells treated with TNF-α (50 ng/ml) or PBS were amplified by PCR. ( E ) The promoter activity was evaluated by transfection with different CCR4 luciferase expression vectors. Cells were treated with TNF-α (50 ng/ml) or PBS for 12 h.

Article Snippet: pGLV-GFP-CCR4 lentiviral vector and four shRNA plasmids targeting different regions of CCR4 mRNA were purchased from Genepharma (Shanghai, China).

Techniques: Incubation, Expressing, Western Blot, Immunohistochemistry, Microarray, Amplification, Activity Assay, Transfection, Luciferase

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: TOE1 is an inhibitor of HIV-1 replication with cell-penetrating capability

doi: 10.1073/pnas.1500857112

Figure Lengend Snippet: TOE1 is a substrate for GraB. (A) Time course in vitro digestion of recombinant TOE1 with 100 nM recombinant active GraB. Cleavage products of TOE1 are revealed by Western blot analysis using a rabbit polyclonal antibody (GeneTex) targeting a central region of TOE1. Incubation with the GraB inhibitor tetrapeptide IETD-CHO (100 μM) prevents the processing. (Lower) Longer exposure of the Upper panel. (B) Identification of D328, D363/373, and D387 cleavage sites in TOE1 by GraB. Constructs expressing either WT FLAG-tagged TOE1 or recombinant forms carrying point mutations in the indicated aspartate residues were transfected into 293T cells; α-FLAG immunoprecipitates from cell lysates were incubated with 100 nM GraB for 2 h. Rabbit α-TOE1 (Ab-86) antibody was used for Western blotting. (C) Western blot analysis of recombinant TOE1 incubated for 2 h with 50 μL of cleared freeze/thaw extract from NK92 cells shows processing of TOE1 that is prevented by the GraB inhibitor Ac-IETD-CHO (100 μM). (Lower) Longer exposure of the Upper panel. (D) Summary of the location of GraB cleavage sites and the positions of the deadenylation (DEDD), C3H zinc finger, and NLS. The C-terminal epitope position for the Ab-86 antibody is also indicated. The GraB tetrapeptide recognition sites for each cleavage site are indicated in brackets.

Article Snippet: Cleavage products of TOE1 are revealed by Western blot analysis using a rabbit polyclonal antibody (GeneTex) targeting a central region of TOE1.

Techniques: In Vitro, Recombinant, Western Blot, Incubation, Construct, Expressing, Transfection

Journal: Neuron

Article Title: Rewiring of human neurodevelopmental gene regulatory programs by Human Accelerated Regions (HARs)

doi: 10.1016/j.neuron.2021.08.005

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: CaptureMPRA library design, cloning, and transfection Molecular Inversion Probes (MIPs) targeting flanking regions of all HAR sequences were designed using the MIPgen program ( Boyle et al., 2014 ) and manufactured as a pool by CustomArray (Bothell, WA).

Techniques: Virus, Recombinant, Electron Microscopy, Modification, Lysis, Sonication, Plasmid Preparation, Software

Journal: Neuron

Article Title: Rewiring of human neurodevelopmental gene regulatory programs by Human Accelerated Regions (HARs)

doi: 10.1016/j.neuron.2021.08.005

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: CaptureMPRA library design, cloning, and transfection Molecular Inversion Probes (MIPs) targeting flanking regions of all HAR sequences were designed using the MIPgen program ( Boyle et al., 2014 ) and manufactured as a pool by CustomArray (Bothell, WA).

Techniques: Virus, Recombinant, Electron Microscopy, Modification, Lysis, Sonication, Plasmid Preparation, Software