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: Scientific Reports

Article Title: EVI5 unveils a role of the oncogene in modulating the Rab11/PD-L1 pathway in lung adenocarcinoma

doi: 10.1038/s41598-025-24726-w

Figure Lengend Snippet: Correlation of EVI5 mRNA expression with Rab11 and PD-L1 mRNA levels in LUAD. ( a ) EVI5 mRNA is positively correlated with Rab11 mRNA in TCGA database, ( b ) EVI5 mRNA is positively correlated with PD-L1 mRNA in TCGA database. ( c ) Effect of the EVI5 mRNA expression level on overall survival in 1161 LUAD patients. Kaplan–Meier plots were generated using Kaplan–Meier Plotter (http://www.kmplot.com). Significant differences: * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: To establish stable EVI5-overexpressing LUAD cell lines, a 2493-bp fragment of the EVI5 coding sequence was synthesized (GeneChem, Shanghai, China) and subcloned into the PLVX-IRES-Neo vector using EcoRI and XbaI.

Techniques: Expressing, Generated

Journal: Scientific Reports

Article Title: EVI5 unveils a role of the oncogene in modulating the Rab11/PD-L1 pathway in lung adenocarcinoma

doi: 10.1038/s41598-025-24726-w

Figure Lengend Snippet: Rab11 and PD-L1 expression in EVI5-KO A549 cells. (a) EVI5 mRNA expression levels. (b) Rab11 mRNA expression levels. (c) PD-L1 mRNA expression levels. (d) Protein levels of EVI5, Rab11, and PD-L1. β-actin served as the internal control. Data represent mean ± SD of three independent experiments. Significant differences: * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: To establish stable EVI5-overexpressing LUAD cell lines, a 2493-bp fragment of the EVI5 coding sequence was synthesized (GeneChem, Shanghai, China) and subcloned into the PLVX-IRES-Neo vector using EcoRI and XbaI.

Techniques: Expressing, Control

Journal: Scientific Reports

Article Title: EVI5 unveils a role of the oncogene in modulating the Rab11/PD-L1 pathway in lung adenocarcinoma

doi: 10.1038/s41598-025-24726-w

Figure Lengend Snippet: Immunohistochemical analysis of EVI5, Rab11, and PD-L1 expression in LUAD tissues. ( a ) Expression levels of EVI5, Rab11, and PD-L1 were assessed in LUAD tissues and compared to adjacent noncancerous lung tissues. ( b ) EVI5 expression was evaluated by IHC and semi-quantitatively assesed using an H-score methodology. ( c ) Rab11 expression was evaluated by IHC and semi-quantitatively assesed using an H-score methodology. ( d ) PD-L1 expression was evaluated by IHC and semi-quantitatively assesed using an H-score methodology.

Article Snippet: To establish stable EVI5-overexpressing LUAD cell lines, a 2493-bp fragment of the EVI5 coding sequence was synthesized (GeneChem, Shanghai, China) and subcloned into the PLVX-IRES-Neo vector using EcoRI and XbaI.

Techniques: Immunohistochemical staining, Expressing

Journal: Scientific Reports

Article Title: EVI5 unveils a role of the oncogene in modulating the Rab11/PD-L1 pathway in lung adenocarcinoma

doi: 10.1038/s41598-025-24726-w

Figure Lengend Snippet: Rab11 and PD-L1 expression in EVI5-overexpressing A549 cells. ( a ) EVI5 mRNA expression levels. ( b ) Rab11 mRNA expression levels. ( c ) PD-L1 mRNA expression levels. ( d ) Protein levels of EVI5, Rab11, and PD-L1. β-actin served as the internal control. Data represent mean ± SD of three independent experiments. Significant differences: * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: To establish stable EVI5-overexpressing LUAD cell lines, a 2493-bp fragment of the EVI5 coding sequence was synthesized (GeneChem, Shanghai, China) and subcloned into the PLVX-IRES-Neo vector using EcoRI and XbaI.

Techniques: Expressing, Control

Journal: Scientific Reports

Article Title: EVI5 unveils a role of the oncogene in modulating the Rab11/PD-L1 pathway in lung adenocarcinoma

doi: 10.1038/s41598-025-24726-w

Figure Lengend Snippet: Rab11 and PD-L1 expression in both EVI5-overexpression and Rab11-knockdown A549 cells. ( a ) Protein levels of EVI5, Rab11, and PD-L1. ( b ) EVI5 mRNA expression levels. ( c ) Rab11 mRNA expression levels. ( d ) PD-L1 mRNA expression levels. Data represent the mean ± SD of three independent experiments. Significant differences: * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: To establish stable EVI5-overexpressing LUAD cell lines, a 2493-bp fragment of the EVI5 coding sequence was synthesized (GeneChem, Shanghai, China) and subcloned into the PLVX-IRES-Neo vector using EcoRI and XbaI.

Techniques: Expressing, Over Expression, Knockdown

Journal: Scientific Reports

Article Title: EVI5 unveils a role of the oncogene in modulating the Rab11/PD-L1 pathway in lung adenocarcinoma

doi: 10.1038/s41598-025-24726-w

Figure Lengend Snippet: Rab11 and PD-L1 expression in PD-L1 inhibition A549 cells. ( a ) IC50 of PLVX cells in inhibiting PD-L1. ( b ) IC50 of EVI5-OE cells in inhibiting PD-L1. ( c ) Protein levels of EVI5, Rab11, and PD-L1. ( d ) EVI5 mRNA expression levels. ( e ) Rab11 mRNA expression levels. ( f ) PD-L1 mRNA expression levels. Data represent the mean ± SD of three independent experiments. Significant differences: * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: To establish stable EVI5-overexpressing LUAD cell lines, a 2493-bp fragment of the EVI5 coding sequence was synthesized (GeneChem, Shanghai, China) and subcloned into the PLVX-IRES-Neo vector using EcoRI and XbaI.

Techniques: Expressing, Inhibition

Journal: Scientific Reports

Article Title: EVI5 unveils a role of the oncogene in modulating the Rab11/PD-L1 pathway in lung adenocarcinoma

doi: 10.1038/s41598-025-24726-w

Figure Lengend Snippet: EVI5, Rab11, and PD-L1 Are Highly Correlated in LUAD Cells. ( a ) Immunofluorescence staining showing co-expression of PD-L1 and Rab11 in EVI5-overexpressing cells compared to control cells (scale bar: 5 μm). ( b ) Analysis of PD-L1and Rab11 co-located Mander coefficients. M1 represents the proportion of co-localization in the red channel, and M2 represents the proportion of co-localization in the green channel. ( c ) Immunofluorescence staining showing co-expression of PD-L1 and EVI5 in EVI5-overexpressing cells compared to control cells (scale bar: 5 μm). ( d ) Analysis of PD-L1and EVI5 co-located Mander coefficients. M1 represents the proportion of co-localization in the red channel, and M2 represents the proportion of co-localization in the green channel. ( e ) Immunofluorescence staining showing co-expression of EVI5 and Rab11 in EVI5-overexpressing cells compared to control cells (scale bar: 5 μm). ( f ) Analysis of Rab11 and EVI5 co-located Mander coefficients. M1 represents the proportion of co-localization in the red channel, and M2 represents the proportion of co-localization in the green channel.

Article Snippet: To establish stable EVI5-overexpressing LUAD cell lines, a 2493-bp fragment of the EVI5 coding sequence was synthesized (GeneChem, Shanghai, China) and subcloned into the PLVX-IRES-Neo vector using EcoRI and XbaI.

Techniques: Immunofluorescence, Staining, Expressing, Control

Journal: Scientific Reports

Article Title: EVI5 unveils a role of the oncogene in modulating the Rab11/PD-L1 pathway in lung adenocarcinoma

doi: 10.1038/s41598-025-24726-w

Figure Lengend Snippet: A mechanistic working model of EVI5 in controlling Rab11/PD-L1 signaling pathway: EVI5 promotes tumor progression through modulation of the Rab11/PD-L1 signaling axis in LUAD.

Article Snippet: To establish stable EVI5-overexpressing LUAD cell lines, a 2493-bp fragment of the EVI5 coding sequence was synthesized (GeneChem, Shanghai, China) and subcloned into the PLVX-IRES-Neo vector using EcoRI and XbaI.

Techniques: