Journal: Frontiers in Oncology

Article Title: A Cell Cycle-Related 13-mRNA Signature to Predict Prognosis in Hepatocellular Carcinoma

doi: 10.3389/fonc.2022.760190

Figure Lengend Snippet: Expression verification and functional experiments. (A) mRNA levels of the 13 genes in the HCC samples (n = 50) compared to normal peritumoral tissues (n = 50) in TCGA cohort. Expression of (B) SPDL1, (C) BRSK1, and (D) TICRR in HCC tissues (n = 23) compared to normal peritumoral tissues (n = 23), as measured by RT-qPCR. (E) Expression of TICRR in HUH-7, Hep3B, and HepG2 cells, as measured by RT-qPCR. (F) Knockdown efficiency of TICRR in Hep3B and HepG2 using three independent siRNAs. (G) CCK-8 analysis showed that depletion of TICRR in Hep3B and HepG2 cells by 2 separate siRNAs led to decreased cell proliferation. *P < 0.05, **P < 0.01, and ***P < 0.001. NS, nonsignificant.

Article Snippet: Each reaction was conducted with 10-μl mixture containing SYBR Green qPCR Master Mix (MCE; No.: HY-K0501), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used for normalization.

Techniques: Expressing, Functional Assay, Quantitative RT-PCR, Knockdown, CCK-8 Assay

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

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

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

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

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

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

Journal: Frontiers in Immunology

Article Title: Inhibition of IL1R1 or CASP4 attenuates spinal cord injury through ameliorating NLRP3 inflammasome-induced pyroptosis

doi: 10.3389/fimmu.2022.963582

Figure Lengend Snippet: Primer sequences for RT-qPCR.

Article Snippet: Next, RT-qPCR was performed using AceQ qPCR SYBR Green Master Mix (Q111-02, Vazyme, China) in a 7500 real-time PCR system (Applied Biosystems, Inc., USA) according to the manufacturer’s instruction.

Techniques:

Journal: Frontiers in Immunology

Article Title: Inhibition of IL1R1 or CASP4 attenuates spinal cord injury through ameliorating NLRP3 inflammasome-induced pyroptosis

doi: 10.3389/fimmu.2022.963582

Figure Lengend Snippet: Western blot, RT-qPCR and immunohistochemistry of the three biomarkers (IL1R1, CASP4 and IGSF6). (A) The time curves of the three biomarkers at mRNA level on day 1, 3, 5 and 7 after SCI or sham surgery (n = 3 rats per group at each time point, values are the mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, t-test). (B) RT-qPCR validation of the biomarkers expressing in peak: the three signatures were increased significantly in SCI group (n = 3 rats per group, values are the mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, t-test). (C) Western blot validation of the biomarkers as represented by their time curves at protein level (n = 3 rats per group, values are the mean ± SD, *p < 0.05, t-test). (D) Histomorphologically, the injured spinal cord tissue exhibited organizational abnormalities as compared with the normal control (overall scale bars = 200μm, regional-scale bars = 100μm). (E) Immunohistochemical validation of the biomarkers: the markers were found to be expressed in both gray matter and white matter (Overall scale bars = 200μm, regional-scale bars = 100μm, the arrows point to the positioning of the marker).

Article Snippet: Next, RT-qPCR was performed using AceQ qPCR SYBR Green Master Mix (Q111-02, Vazyme, China) in a 7500 real-time PCR system (Applied Biosystems, Inc., USA) according to the manufacturer’s instruction.

Techniques: Western Blot, Quantitative RT-PCR, Immunohistochemistry, Biomarker Discovery, Expressing, Control, Immunohistochemical staining, Marker

Journal: Frontiers in Immunology

Article Title: Inhibition of IL1R1 or CASP4 attenuates spinal cord injury through ameliorating NLRP3 inflammasome-induced pyroptosis

doi: 10.3389/fimmu.2022.963582

Figure Lengend Snippet: Change of the biomarkers and functional recovery in the SCI model after immunosuppressive therapy. (A) Statistical analysis of the BBB Scale in Sham, positive control, and treatment groups over 28 days (n = 3 rats per group at each time point, values are the mean ± SD, *p < 0.05, t-test). (B) Footprint analysis of different groups. (C, D) Statistical analysis of the Louisville Swim Scale in the four groups over 28 days (n = 3 rats per group at each time point, values are the mean ± SD, *p < 0.05, t-test). (E) Western blot showed that the expression level of the biomarkers in the treatment groups was between the levels of sham and control groups, and the levels of Bcl2 and CASP3, NFκB, GSDMD, IL1B, ASC, and CASP3 in SA and SV groups were also between those of Sham and control groups (n = 3 rats per group, *p < 0.05, **p < 0.01, ***p < 0.001, t-test). (F) RT-qPCR showed that the level of biomarkers was significantly decreased after treatment (n = 3 rats per group, *p < 0.05, **p < 0.01, ***p < 0.001, t-test).

Article Snippet: Next, RT-qPCR was performed using AceQ qPCR SYBR Green Master Mix (Q111-02, Vazyme, China) in a 7500 real-time PCR system (Applied Biosystems, Inc., USA) according to the manufacturer’s instruction.

Techniques: Functional Assay, Positive Control, Western Blot, Expressing, Control, Quantitative RT-PCR