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IFN-γ treatment enhances epithelial-mesenchymal transition (EMT) and metastasis in various tumors. (A) Transwell assays to detect the migration and invasion abilities of different cancer cell lines without and with IFN-γ treatment (50 ng/ml, 48 h). (B) Western blotting assays to examine the effect of IFN-γ treatment (50 ng/ml, 48 h) on expression of EMT markers at the protein level. (C) <t>RT-qPCR</t> assays to detect the changes in expression of EMT markers at the mRNA level after IFN-γ treatment (50 ng/ml, 48 h). Data are shown as the mean ± standard deviation (SD). *P < 0.05, **P < 0.01, ****P < 0.0001.
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IFN-γ treatment enhances epithelial-mesenchymal transition (EMT) and metastasis in various tumors. (A) Transwell assays to detect the migration and invasion abilities of different cancer cell lines without and with IFN-γ treatment (50 ng/ml, 48 h). (B) Western blotting assays to examine the effect of IFN-γ treatment (50 ng/ml, 48 h) on expression of EMT markers at the protein level. (C) <t>RT-qPCR</t> assays to detect the changes in expression of EMT markers at the mRNA level after IFN-γ treatment (50 ng/ml, 48 h). Data are shown as the mean ± standard deviation (SD). *P < 0.05, **P < 0.01, ****P < 0.0001.
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IFN-γ treatment enhances epithelial-mesenchymal transition (EMT) and metastasis in various tumors. (A) Transwell assays to detect the migration and invasion abilities of different cancer cell lines without and with IFN-γ treatment (50 ng/ml, 48 h). (B) Western blotting assays to examine the effect of IFN-γ treatment (50 ng/ml, 48 h) on expression of EMT markers at the protein level. (C) <t>RT-qPCR</t> assays to detect the changes in expression of EMT markers at the mRNA level after IFN-γ treatment (50 ng/ml, 48 h). Data are shown as the mean ± standard deviation (SD). *P < 0.05, **P < 0.01, ****P < 0.0001.
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IFN-γ treatment enhances epithelial-mesenchymal transition (EMT) and metastasis in various tumors. (A) Transwell assays to detect the migration and invasion abilities of different cancer cell lines without and with IFN-γ treatment (50 ng/ml, 48 h). (B) Western blotting assays to examine the effect of IFN-γ treatment (50 ng/ml, 48 h) on expression of EMT markers at the protein level. (C) <t>RT-qPCR</t> assays to detect the changes in expression of EMT markers at the mRNA level after IFN-γ treatment (50 ng/ml, 48 h). Data are shown as the mean ± standard deviation (SD). *P < 0.05, **P < 0.01, ****P < 0.0001.
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IFN-γ treatment enhances epithelial-mesenchymal transition (EMT) and metastasis in various tumors. (A) Transwell assays to detect the migration and invasion abilities of different cancer cell lines without and with IFN-γ treatment (50 ng/ml, 48 h). (B) Western blotting assays to examine the effect of IFN-γ treatment (50 ng/ml, 48 h) on expression of EMT markers at the protein level. (C) <t>RT-qPCR</t> assays to detect the changes in expression of EMT markers at the mRNA level after IFN-γ treatment (50 ng/ml, 48 h). Data are shown as the mean ± standard deviation (SD). *P < 0.05, **P < 0.01, ****P < 0.0001.
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IFN-γ treatment enhances epithelial-mesenchymal transition (EMT) and metastasis in various tumors. (A) Transwell assays to detect the migration and invasion abilities of different cancer cell lines without and with IFN-γ treatment (50 ng/ml, 48 h). (B) Western blotting assays to examine the effect of IFN-γ treatment (50 ng/ml, 48 h) on expression of EMT markers at the protein level. (C) <t>RT-qPCR</t> assays to detect the changes in expression of EMT markers at the mRNA level after IFN-γ treatment (50 ng/ml, 48 h). Data are shown as the mean ± standard deviation (SD). *P < 0.05, **P < 0.01, ****P < 0.0001.
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IFN-γ treatment enhances epithelial-mesenchymal transition (EMT) and metastasis in various tumors. (A) Transwell assays to detect the migration and invasion abilities of different cancer cell lines without and with IFN-γ treatment (50 ng/ml, 48 h). (B) Western blotting assays to examine the effect of IFN-γ treatment (50 ng/ml, 48 h) on expression of EMT markers at the protein level. (C) <t>RT-qPCR</t> assays to detect the changes in expression of EMT markers at the mRNA level after IFN-γ treatment (50 ng/ml, 48 h). Data are shown as the mean ± standard deviation (SD). *P < 0.05, **P < 0.01, ****P < 0.0001.
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IFN-γ treatment enhances epithelial-mesenchymal transition (EMT) and metastasis in various tumors. (A) Transwell assays to detect the migration and invasion abilities of different cancer cell lines without and with IFN-γ treatment (50 ng/ml, 48 h). (B) Western blotting assays to examine the effect of IFN-γ treatment (50 ng/ml, 48 h) on expression of EMT markers at the protein level. (C) <t>RT-qPCR</t> assays to detect the changes in expression of EMT markers at the mRNA level after IFN-γ treatment (50 ng/ml, 48 h). Data are shown as the mean ± standard deviation (SD). *P < 0.05, **P < 0.01, ****P < 0.0001.
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IFN-γ treatment enhances epithelial-mesenchymal transition (EMT) and metastasis in various tumors. (A) Transwell assays to detect the migration and invasion abilities of different cancer cell lines without and with IFN-γ treatment (50 ng/ml, 48 h). (B) Western blotting assays to examine the effect of IFN-γ treatment (50 ng/ml, 48 h) on expression of EMT markers at the protein level. (C) <t>RT-qPCR</t> assays to detect the changes in expression of EMT markers at the mRNA level after IFN-γ treatment (50 ng/ml, 48 h). Data are shown as the mean ± standard deviation (SD). *P < 0.05, **P < 0.01, ****P < 0.0001.
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IFN-γ treatment enhances epithelial-mesenchymal transition (EMT) and metastasis in various tumors. (A) Transwell assays to detect the migration and invasion abilities of different cancer cell lines without and with IFN-γ treatment (50 ng/ml, 48 h). (B) Western blotting assays to examine the effect of IFN-γ treatment (50 ng/ml, 48 h) on expression of EMT markers at the protein level. (C) <t>RT-qPCR</t> assays to detect the changes in expression of EMT markers at the mRNA level after IFN-γ treatment (50 ng/ml, 48 h). Data are shown as the mean ± standard deviation (SD). *P < 0.05, **P < 0.01, ****P < 0.0001.
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IFN-γ treatment enhances epithelial-mesenchymal transition (EMT) and metastasis in various tumors. (A) Transwell assays to detect the migration and invasion abilities of different cancer cell lines without and with IFN-γ treatment (50 ng/ml, 48 h). (B) Western blotting assays to examine the effect of IFN-γ treatment (50 ng/ml, 48 h) on expression of EMT markers at the protein level. (C) RT-qPCR assays to detect the changes in expression of EMT markers at the mRNA level after IFN-γ treatment (50 ng/ml, 48 h). Data are shown as the mean ± standard deviation (SD). *P < 0.05, **P < 0.01, ****P < 0.0001.

Journal: Frontiers in Oncology

Article Title: Targeting MDK Abrogates IFN-γ-Elicited Metastasis inCancers of Various Origins

doi: 10.3389/fonc.2022.885656

Figure Lengend Snippet: IFN-γ treatment enhances epithelial-mesenchymal transition (EMT) and metastasis in various tumors. (A) Transwell assays to detect the migration and invasion abilities of different cancer cell lines without and with IFN-γ treatment (50 ng/ml, 48 h). (B) Western blotting assays to examine the effect of IFN-γ treatment (50 ng/ml, 48 h) on expression of EMT markers at the protein level. (C) RT-qPCR assays to detect the changes in expression of EMT markers at the mRNA level after IFN-γ treatment (50 ng/ml, 48 h). Data are shown as the mean ± standard deviation (SD). *P < 0.05, **P < 0.01, ****P < 0.0001.

Article Snippet: QRT-PCR was performed using qPCR SYBR Green Master Mix (Yeasen Biotechnology, Shanghai, China).

Techniques: Migration, Western Blot, Expressing, Quantitative RT-PCR, Standard Deviation

IFN-γ treatment promotes MDK expression in different cancers. (A) RT-qPCR assays to detect the expression of MDK in five cancer cell lines without and with IFN-γ treatment (50 ng/ml, 48 h). (B) Western blotting assay to determine the influence of IFN-γ treatment (50 ng/ml, 48 h) on MDK protein levels in five cancer cell lines. Data are shown as the mean ± standard deviation (SD). **P < 0.01, ***P < 0.001, ****P < 0.0001.

Journal: Frontiers in Oncology

Article Title: Targeting MDK Abrogates IFN-γ-Elicited Metastasis inCancers of Various Origins

doi: 10.3389/fonc.2022.885656

Figure Lengend Snippet: IFN-γ treatment promotes MDK expression in different cancers. (A) RT-qPCR assays to detect the expression of MDK in five cancer cell lines without and with IFN-γ treatment (50 ng/ml, 48 h). (B) Western blotting assay to determine the influence of IFN-γ treatment (50 ng/ml, 48 h) on MDK protein levels in five cancer cell lines. Data are shown as the mean ± standard deviation (SD). **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: QRT-PCR was performed using qPCR SYBR Green Master Mix (Yeasen Biotechnology, Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation

Dose- and time-dependent effects of IFN-γ on MDK induction. (A) RT-qPCR assays to examine the effect of IFN-γ concentration on MDK induction in the HCT116 cell line. (B) Western blotting assays to examine the effect of IFN-γ concentration on MDK induction in the HCT116 cell line. (C) RT-qPCR assays to examine the effect of IFN-γ treatment time on MDK induction in the HCT116 cell line. (D) Western blotting assays to examine the effect of IFN-γ treatment time on MDK induction and EMT activation in the HCT116 cell line. (E) RT-qPCR assays showing the effect of IFN-γ treatment time on EMT markers in the HCT116 cell line. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. ns, not significant.

Journal: Frontiers in Oncology

Article Title: Targeting MDK Abrogates IFN-γ-Elicited Metastasis inCancers of Various Origins

doi: 10.3389/fonc.2022.885656

Figure Lengend Snippet: Dose- and time-dependent effects of IFN-γ on MDK induction. (A) RT-qPCR assays to examine the effect of IFN-γ concentration on MDK induction in the HCT116 cell line. (B) Western blotting assays to examine the effect of IFN-γ concentration on MDK induction in the HCT116 cell line. (C) RT-qPCR assays to examine the effect of IFN-γ treatment time on MDK induction in the HCT116 cell line. (D) Western blotting assays to examine the effect of IFN-γ treatment time on MDK induction and EMT activation in the HCT116 cell line. (E) RT-qPCR assays showing the effect of IFN-γ treatment time on EMT markers in the HCT116 cell line. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. ns, not significant.

Article Snippet: QRT-PCR was performed using qPCR SYBR Green Master Mix (Yeasen Biotechnology, Shanghai, China).

Techniques: Quantitative RT-PCR, Concentration Assay, Western Blot, Activation Assay

IFN-γ activates MDK via STAT1. (A) Correlation analyses using the GEPIA 2 online tool ( http://gepia2.cancer-pku.cn/#correlation ) to shown the correlation relationship between STAT1 and MDK using the RNA-seq data of different cancers from the Cancer Genome Atlas (TCGA) database. KIRC, kidney renal clear cell carcinoma; LUSC, Lung squamous cell carcinoma; CESC, Cervical squamous cell carcinoma and endocervical adenocarcinoma; BRCA, breast invasive carcinoma; COAD, colorectal adenocarcinoma. (B) RT-qPCR and Western blotting assays to detect the effect of IFN-γ treatment (50 ng/ml, 48h) on levels of STAT1 and phosphorylated STAT1 (p-STAT1) in five cancer cell lines. (C) RT-qPCR assays to the effect of STAT1 inhibitor on mRNA levels of STAT1 and MDK in five cancer cell lines. (D) Western blotting assays to examine the effect of STAT1 inhibitor on protein levels of STAT1, p-STAT1 and MDK in five cancer cell lines. Data are shown as the mean ± standard deviation (SD). **P < 0.01, ***P < 0.001, ****P < 0.0001.

Journal: Frontiers in Oncology

Article Title: Targeting MDK Abrogates IFN-γ-Elicited Metastasis inCancers of Various Origins

doi: 10.3389/fonc.2022.885656

Figure Lengend Snippet: IFN-γ activates MDK via STAT1. (A) Correlation analyses using the GEPIA 2 online tool ( http://gepia2.cancer-pku.cn/#correlation ) to shown the correlation relationship between STAT1 and MDK using the RNA-seq data of different cancers from the Cancer Genome Atlas (TCGA) database. KIRC, kidney renal clear cell carcinoma; LUSC, Lung squamous cell carcinoma; CESC, Cervical squamous cell carcinoma and endocervical adenocarcinoma; BRCA, breast invasive carcinoma; COAD, colorectal adenocarcinoma. (B) RT-qPCR and Western blotting assays to detect the effect of IFN-γ treatment (50 ng/ml, 48h) on levels of STAT1 and phosphorylated STAT1 (p-STAT1) in five cancer cell lines. (C) RT-qPCR assays to the effect of STAT1 inhibitor on mRNA levels of STAT1 and MDK in five cancer cell lines. (D) Western blotting assays to examine the effect of STAT1 inhibitor on protein levels of STAT1, p-STAT1 and MDK in five cancer cell lines. Data are shown as the mean ± standard deviation (SD). **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: QRT-PCR was performed using qPCR SYBR Green Master Mix (Yeasen Biotechnology, Shanghai, China).

Techniques: RNA Sequencing, Quantitative RT-PCR, Western Blot, Standard Deviation

MDK promotes cancer metastasis by activating the EMT programme. (A) RT-qPCR validation of MDK overexpression in five cancer cell lines. (B) Transwell assays to assess the alteration of cell invasion and migration abilities by MDK overexpression in five cancer cell lines. (C) Western blotting assays to assess the effect of MDK overexpression on expression of EMT markers. (D) RT-qPCR assays to assess the effect of MDK overexpression on expression of EMT markers. (E) RT-qPCR validation of MDK inhibition by iMDK (100nM, 48h). (F) RT-qPCR and Western blotting assays to assess the effect of iMDK on EMT in five cancer cell lines. (G) Transwell assays to assess the effect of iMDK on cell invasion and migration in five cancer cell lines. Data are shown as the mean ± standard deviation (SD). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Journal: Frontiers in Oncology

Article Title: Targeting MDK Abrogates IFN-γ-Elicited Metastasis inCancers of Various Origins

doi: 10.3389/fonc.2022.885656

Figure Lengend Snippet: MDK promotes cancer metastasis by activating the EMT programme. (A) RT-qPCR validation of MDK overexpression in five cancer cell lines. (B) Transwell assays to assess the alteration of cell invasion and migration abilities by MDK overexpression in five cancer cell lines. (C) Western blotting assays to assess the effect of MDK overexpression on expression of EMT markers. (D) RT-qPCR assays to assess the effect of MDK overexpression on expression of EMT markers. (E) RT-qPCR validation of MDK inhibition by iMDK (100nM, 48h). (F) RT-qPCR and Western blotting assays to assess the effect of iMDK on EMT in five cancer cell lines. (G) Transwell assays to assess the effect of iMDK on cell invasion and migration in five cancer cell lines. Data are shown as the mean ± standard deviation (SD). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: QRT-PCR was performed using qPCR SYBR Green Master Mix (Yeasen Biotechnology, Shanghai, China).

Techniques: Quantitative RT-PCR, Biomarker Discovery, Over Expression, Migration, Western Blot, Expressing, Inhibition, Standard Deviation

Targeting MDK abrogates IFN-γ-induced tumor metastasis. (A) RT-qPCR validation of MDK suppression by MDK inhibitor in IFN-γ-treated cancer cells. (B) Western blotting and RT-qPCR assays to assess the effect of iMDK on IFN-γ-activated EMT in five cancer cell lines. (C) Transwell assays to examine the effect of iMDK on IFN-γ-driven cell invasion and migration. Data are shown as the mean ± standard deviation (SD). **P < 0.01, ***P < 0.001, ****P < 0.0001.

Journal: Frontiers in Oncology

Article Title: Targeting MDK Abrogates IFN-γ-Elicited Metastasis inCancers of Various Origins

doi: 10.3389/fonc.2022.885656

Figure Lengend Snippet: Targeting MDK abrogates IFN-γ-induced tumor metastasis. (A) RT-qPCR validation of MDK suppression by MDK inhibitor in IFN-γ-treated cancer cells. (B) Western blotting and RT-qPCR assays to assess the effect of iMDK on IFN-γ-activated EMT in five cancer cell lines. (C) Transwell assays to examine the effect of iMDK on IFN-γ-driven cell invasion and migration. Data are shown as the mean ± standard deviation (SD). **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: QRT-PCR was performed using qPCR SYBR Green Master Mix (Yeasen Biotechnology, Shanghai, China).

Techniques: Quantitative RT-PCR, Biomarker Discovery, Western Blot, Migration, Standard Deviation