Journal: Scientific Reports

Article Title: Thermal transport analysis of six circular microchannel heat sink using nanofluid

doi: 10.1038/s41598-022-11121-y

Figure Lengend Snippet: Properties of microchannel heat sink dimensions.

Article Snippet: Figure 19 Temperature contour for a microelectronic chip in a microchannel heat sink with a nanofluid utilizing ANSYS.

Techniques:

Journal: Scientific Reports

Article Title: Thermal transport analysis of six circular microchannel heat sink using nanofluid

doi: 10.1038/s41598-022-11121-y

Figure Lengend Snippet: Computational mesh of the micropolar chip for cooling chip: ( a ) side view along the xy-plane, ( b ) bottom view, and ( c ) microchannel.

Article Snippet: Figure 19 Temperature contour for a microelectronic chip in a microchannel heat sink with a nanofluid utilizing ANSYS.

Techniques:

Journal: Scientific Reports

Article Title: Thermal transport analysis of six circular microchannel heat sink using nanofluid

doi: 10.1038/s41598-022-11121-y

Figure Lengend Snippet: Estimation of the Nusselt number against inlet fluid velocity across the microchannel for an electronic chip heat sink with \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\text{TiO}}_{2}$$\end{document} TiO 2 /water nanofluid and distilled water.

Article Snippet: Figure 19 Temperature contour for a microelectronic chip in a microchannel heat sink with a nanofluid utilizing ANSYS.

Techniques:

Journal: Scientific Reports

Article Title: Thermal transport analysis of six circular microchannel heat sink using nanofluid

doi: 10.1038/s41598-022-11121-y

Figure Lengend Snippet: Estimation of the friction factor against the Reynolds number in a microchannel heat sink with \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\text{TiO}}_{2}$$\end{document} TiO 2 /water nanofluids and distilled water.

Article Snippet: Figure 19 Temperature contour for a microelectronic chip in a microchannel heat sink with a nanofluid utilizing ANSYS.

Techniques:

Journal: Scientific Reports

Article Title: Thermal transport analysis of six circular microchannel heat sink using nanofluid

doi: 10.1038/s41598-022-11121-y

Figure Lengend Snippet: Estimation of the Wall temperature against the Reynolds number in a microchannel heat sink with \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${\text{TiO}}_{2}$$\end{document} TiO 2 /water nanofluid and distilled water.

Article Snippet: Figure 19 Temperature contour for a microelectronic chip in a microchannel heat sink with a nanofluid utilizing ANSYS.

Techniques:

Journal: Scientific Reports

Article Title: Thermal transport analysis of six circular microchannel heat sink using nanofluid

doi: 10.1038/s41598-022-11121-y

Figure Lengend Snippet: Temperature contour for a microelectronic chip in a microchannel heat sink with a nanofluid utilizing ANSYS.

Article Snippet: Figure 19 Temperature contour for a microelectronic chip in a microchannel heat sink with a nanofluid utilizing ANSYS.

Techniques:

Journal: Scientific Reports

Article Title: Thermal transport analysis of six circular microchannel heat sink using nanofluid

doi: 10.1038/s41598-022-11121-y

Figure Lengend Snippet: Wall heat flux significance for microelectronic chips in microchannel heat sinks with nanofluids by using ANSYS.

Article Snippet: Figure 19 Temperature contour for a microelectronic chip in a microchannel heat sink with a nanofluid utilizing ANSYS.

Techniques:

Journal: Scientific Reports

Article Title: Thermal transport analysis of six circular microchannel heat sink using nanofluid

doi: 10.1038/s41598-022-11121-y

Figure Lengend Snippet: Cross-sectional Schmidt of the wall heat flux contour in the microchannel heat sink .

Article Snippet: Figure 19 Temperature contour for a microelectronic chip in a microchannel heat sink with a nanofluid utilizing ANSYS.

Techniques:

Journal: Cell death & disease

Article Title: TRIB3 promotes malignancy of head and neck squamous cell carcinoma via inhibiting ferroptosis.

doi: 10.1038/s41419-024-06472-5

Figure Lengend Snippet: Fig. 1 Associations between prognostic significance and TRIB3 expression in HNSCC. A A Heat Map of TRIB1, TRIB2, and TRIB3 gene expression in HNSCC and normal tissues. B, C Comparison analysis of TRIB3 levels among HNSCC tissues and normal tissues or adjacent tissues in the TCGA database or the CPTAC database. D ROC curve indicating the predictive value of TRIB3. E Comparison of the low and high expression of TRIB3 via Kaplan–Meier OS curve and survival status in GEO database or F TCGA database. G Comparison between the low and high expression of TRIB3 via Kaplan–Meier PFS curve and survival status in TCGA database. ***P < 0.001, **P < 0.01, *P < 0.05.

Article Snippet: Following are the antibodies utilized for immunofluorescence and their concentrations: TRIB3 (Proteintech, China, 13300-1-AP, 1:100), β-catenin (Cell Signaling Technology, 2677 S, 1:200), TCF4 (NOVUS, H00006925-M03, 10 μg/ml), Anti-rat IgG (abcam, ab15007, 1:1,000), Anti-mouse IgG (abcam, ab150115, 1:1,000).

Techniques: Expressing, Gene Expression, Comparison

Journal: Cell death & disease

Article Title: TRIB3 promotes malignancy of head and neck squamous cell carcinoma via inhibiting ferroptosis.

doi: 10.1038/s41419-024-06472-5

Figure Lengend Snippet: Fig. 2 TRIB3 silencing inhibits HNSCC cell viability and proliferation. A Western blot to detect protein levels of TRIB3 in normal NOK cell and HNSCC cell lines. Cal27 and FaDu cells were transfected with two independent TRIB3 shRNAs. Western blot (B) and qPCR (C) to detect TRIB3 levels in TRIB3-knockdown and control cells. D OD450 readings were plotted over time using the CCK8 assay. E, F Findings of a clonogenic analysis. Red circles demonstrated the counted clones. G Visualizztion of DNA replication using EdU. Red-stained cell nuclei demonstrated DNA replication. Scale bar: 50 μm. ***P < 0.001, **P < 0.01, *P < 0.05.

Article Snippet: Following are the antibodies utilized for immunofluorescence and their concentrations: TRIB3 (Proteintech, China, 13300-1-AP, 1:100), β-catenin (Cell Signaling Technology, 2677 S, 1:200), TCF4 (NOVUS, H00006925-M03, 10 μg/ml), Anti-rat IgG (abcam, ab15007, 1:1,000), Anti-mouse IgG (abcam, ab150115, 1:1,000).

Techniques: Western Blot, Transfection, Knockdown, Control, CCK-8 Assay, Clone Assay, Staining

Journal: Cell death & disease

Article Title: TRIB3 promotes malignancy of head and neck squamous cell carcinoma via inhibiting ferroptosis.

doi: 10.1038/s41419-024-06472-5

Figure Lengend Snippet: Fig. 3 Promotion of cell death by TRIB3 silencing largely through inhibiting ferroptosis. A GO analysis of DEGs between Cal27 TRIB3- knockdown and control cells. B CCK-8 assay detected the cell viability of Cal27 TRIB3-knockdown and control cells, treated with or without cell death inhibitors. C, D Lipid peroxidation of TRIB3-knockdown and control cells. E MDA in TRIB3-knockdown and control cells. F The Fe2+ level in control cells and TRIB3-knockdown. G Transmission electron microscopy images of Cal27 TRIB3-knockdown and control cells. The red arrows demonstrate mitochondria. Yellow arrows demonstrate autophagosomes. Scale bars: right, 500 nm; left, 1 µm. H qPCR analysis of CHAC1 expression in control and TRIB3-knockdown cells. ***P < 0.001, **P < 0.01, *P < 0.05.

Article Snippet: Following are the antibodies utilized for immunofluorescence and their concentrations: TRIB3 (Proteintech, China, 13300-1-AP, 1:100), β-catenin (Cell Signaling Technology, 2677 S, 1:200), TCF4 (NOVUS, H00006925-M03, 10 μg/ml), Anti-rat IgG (abcam, ab15007, 1:1,000), Anti-mouse IgG (abcam, ab150115, 1:1,000).

Techniques: Knockdown, Control, CCK-8 Assay, Transmission Assay, Electron Microscopy, Expressing

Journal: Cell death & disease

Article Title: TRIB3 promotes malignancy of head and neck squamous cell carcinoma via inhibiting ferroptosis.

doi: 10.1038/s41419-024-06472-5

Figure Lengend Snippet: Fig. 4 Induction of ferroptosis by TRIB3 silencing in HNSCC. A, B MDA in control and TRIB3-knockdown cells treated with Ferrostatin-1 (5 μm) or DFO (50 μm) for 48 h. C, D, E Lipid peroxidation in control and TRIB3-knockdown cells treated with Ferrostatin-1 (5 μm) or DFO (50 μm) for 48 h. F, G, H The level of Fe2+ in control and TRIB3-knockdown cells treated with Ferrostatin-1 (5 μm) or DFO (50 μm) for 48 h. I Schematic of TRIB3-knockdown subcutaneous tumors either treated with or without Liproxstatin-1. J, K Tumor growth and L weight in the xenograft model. M The expression of Ki67, TRIB3, 4-HNE using IHC assay. Scale bar: 100 μm. ***P < 0.001, **P < 0.01, *P < 0.05.

Article Snippet: Following are the antibodies utilized for immunofluorescence and their concentrations: TRIB3 (Proteintech, China, 13300-1-AP, 1:100), β-catenin (Cell Signaling Technology, 2677 S, 1:200), TCF4 (NOVUS, H00006925-M03, 10 μg/ml), Anti-rat IgG (abcam, ab15007, 1:1,000), Anti-mouse IgG (abcam, ab150115, 1:1,000).

Techniques: Control, Knockdown, Expressing

Journal: Cell death & disease

Article Title: TRIB3 promotes malignancy of head and neck squamous cell carcinoma via inhibiting ferroptosis.

doi: 10.1038/s41419-024-06472-5

Figure Lengend Snippet: Fig. 5 Interaction of TRIB3 with β-catenin and TCF4 to create a heterotrimeric complex. A KEGG analysis based on RNA-seq showed enrichment of DEGs in Wnt-signaling pathways. B Western blot to detect TCF4, β-catenin, and TRIB3 levels in the cytoplasm and nucleus of control and TRIB3-knockdown cells. C, D HNSCC cells extracts were immuno-precipitated and blotted with anti-TCF4, anti-TRIB3, or anti–β-catenin antibodies, respectively. Normal rabbit IgG was utilized as the control. E, F Co-IP of β-catenin and TCF4 in HNSCC cells with or without TRIB3 knockdown. ***P < 0.001, **P < 0.01, *P < 0.05.

Article Snippet: Following are the antibodies utilized for immunofluorescence and their concentrations: TRIB3 (Proteintech, China, 13300-1-AP, 1:100), β-catenin (Cell Signaling Technology, 2677 S, 1:200), TCF4 (NOVUS, H00006925-M03, 10 μg/ml), Anti-rat IgG (abcam, ab15007, 1:1,000), Anti-mouse IgG (abcam, ab150115, 1:1,000).

Techniques: RNA Sequencing, Protein-Protein interactions, Western Blot, Control, Knockdown, Co-Immunoprecipitation Assay

Journal: Cell death & disease

Article Title: TRIB3 promotes malignancy of head and neck squamous cell carcinoma via inhibiting ferroptosis.

doi: 10.1038/s41419-024-06472-5

Figure Lengend Snippet: Fig. 6 Inhibition of ALOXE3 activity by TRIB3‒β-catenin‒TCF4 heterotrimer complex. A Scatter plot of DEGs. B Overlay of downregulated genes (TRIB3-knockdown vs. control) with known suppressor genes in ferroptosis and upregulated genes (TRIB3-knockdown vs. control) with known driver genes in ferroptosis (up), respectively. Overlay of 12 down DEGs and 6 up DEGs with Cistrome Data Browser ChIP-seq (bottom). C, D qPCR to detect ALOXE3 and PARP8 levels in TRIB3 knockdown and control cells. E, F ChIP analyses of TCF4 binding on the ALOXE3 and PARP8 promoter in Cal27 TRIB3-overexpression cells. G Schematic illustration of wild-type (Wt) and mutant (Mut) sequences of one putative binding sites of TCF4 on ALOXE3 promoter are shown. H Transcriptional activity of ALOXE3 in Cal27 TCF4-knockdown or overexpression cells calculated by the luciferase reporter system. I Western blot to identify ALOXE3 and TCF4 protein level in Cal27 TCF4-knockdown or overexpression cells. ***P < 0.001, **P < 0.01, *P < 0.05.

Article Snippet: Following are the antibodies utilized for immunofluorescence and their concentrations: TRIB3 (Proteintech, China, 13300-1-AP, 1:100), β-catenin (Cell Signaling Technology, 2677 S, 1:200), TCF4 (NOVUS, H00006925-M03, 10 μg/ml), Anti-rat IgG (abcam, ab15007, 1:1,000), Anti-mouse IgG (abcam, ab150115, 1:1,000).

Techniques: Inhibition, Activity Assay, Knockdown, Control, ChIP-sequencing, Binding Assay, Over Expression, Mutagenesis, Luciferase, Western Blot

Journal: Cell death & disease

Article Title: TRIB3 promotes malignancy of head and neck squamous cell carcinoma via inhibiting ferroptosis.

doi: 10.1038/s41419-024-06472-5

Figure Lengend Snippet: Fig. 7 TRIB3‒β-catenin‒TCF4 heterotrimer complex negatively modulates ferroptosis by targeting ALOXE3. (A) Western blot to identify ALOXE3 protein level in TRIB3-knockdown or overexpression and control cells. (B) Western blot to measure ALOXE3 protein level in ALOXE3- knockdown and control cells. (C, D) Lipid peroxidation in ALOXE3-knockdown and control cells. (E) MDA in ALOXE3-knockdown and control cells. (F, G) Lipid peroxidation in HNSCC cells was transfected with TRIB3 and ALOXE3 shRNA. (H) MDA in HNSCC cells was transfected with TRIB3 and ALOXE3 shRNA. ***P < 0.001, **P < 0.01, *P < 0.05.

Article Snippet: Following are the antibodies utilized for immunofluorescence and their concentrations: TRIB3 (Proteintech, China, 13300-1-AP, 1:100), β-catenin (Cell Signaling Technology, 2677 S, 1:200), TCF4 (NOVUS, H00006925-M03, 10 μg/ml), Anti-rat IgG (abcam, ab15007, 1:1,000), Anti-mouse IgG (abcam, ab150115, 1:1,000).

Techniques: Western Blot, Knockdown, Over Expression, Control, Transfection, shRNA

Journal: Cell death & disease

Article Title: TRIB3 promotes malignancy of head and neck squamous cell carcinoma via inhibiting ferroptosis.

doi: 10.1038/s41419-024-06472-5

Figure Lengend Snippet: Fig. 8 Hesperidin suppresses HNSCC initiation and progression. A The active pocket of TRIB3 protein. B The structure of hesperidin. C Cell viability of HNSCC cells treated with different Hesperidin concentrations. D Western blot to detect TRIB3 protein level in HNSCC cells treated with hesperidin (20 μm) for 48 h. E MDA in HNSCC cells treated with hesperidin (20 μm) for 48 h. F, G Lipid peroxidation in HNSCC cells treated with hesperidin (20 μm) for 48 h. H The level of Fe2+ in HNSCC cells treated with hesperidin (20 μm) for 48 h. I Schematic of FaDu subcutaneous tumors treated with hesperidin, J, K tumor growth and L weight in the xenograft model. M The expression of Ki67 using IHC assay. N H&E staining of the spleen, heart, kidney, lung, and liver tissues. Scale bar: 100 μm. O The schematic diagram illustrates the inhibition of ferroptosis in HNSCC by the TRIB3-β-catenin-TCF4 complex through the suppression of ALOXE3 transcription. ***P < 0.001, **P < 0.01, *P < 0.05.

Article Snippet: Following are the antibodies utilized for immunofluorescence and their concentrations: TRIB3 (Proteintech, China, 13300-1-AP, 1:100), β-catenin (Cell Signaling Technology, 2677 S, 1:200), TCF4 (NOVUS, H00006925-M03, 10 μg/ml), Anti-rat IgG (abcam, ab15007, 1:1,000), Anti-mouse IgG (abcam, ab150115, 1:1,000).

Techniques: Western Blot, Expressing, Staining, Inhibition

Journal: Cancers

Article Title: Enhancer of Zeste Homolog 2 Protects Mucosal Melanoma from Ferroptosis via the KLF14-SLC7A11 Signaling Pathway

doi: 10.3390/cancers16213660

Figure Lengend Snippet: Copy number variation of EZH2 gene in melanoma. ( A ) EZH2 gain status in different cancer types from the cBioPortal database. ( B ) Copy number variation of EZH2 gene in 547 melanoma samples. *** p < 0.001. ( C ) Distribution pie chart of EZH2 copy number ( n = 547). According to the EZH2 copy number, it is divided into four subgroups. Subgroup 1: copy number ≤ 2; subgroup 2: 2 < copy number ≤ 3; subgroup 3: 3 < copy number ≤ 4; subgroup 4: copy number > 4. ( D ) Correlation of EZH2 gain status with its mRNA expression in melanoma samples from the cBioPortal database ( n = 367). * p < 0.05, *** p < 0.001. ( E ) Association of EZH2 copy number gain with expression levels in melanoma subtypes ( n = 183). The left panel represents the percentage of cases with or without EZH2 copy number gain across different expression levels (0, 1, 2, 3). The right panel illustrates the proportion of expression levels in different melanoma subtypes: acral melanoma (AM), mucosal melanoma (MM), and cutaneous melanoma (CM). The staining score for each sample, counting the intensity of the staining, was graded as 0, 1, 2, and 3 (“0” as negative, and “3” as the strongest).

Article Snippet: pCMV3-HA vector containing the human EZH2 coding sequence (HA- EZH2 ) was purchased from Sino Biological (HG11337-CY; Sino Biological, Beijing, China).

Techniques: Expressing, Staining

Journal: Cancers

Article Title: Enhancer of Zeste Homolog 2 Protects Mucosal Melanoma from Ferroptosis via the KLF14-SLC7A11 Signaling Pathway

doi: 10.3390/cancers16213660

Figure Lengend Snippet: EZH2 amplification in melanoma.

Article Snippet: pCMV3-HA vector containing the human EZH2 coding sequence (HA- EZH2 ) was purchased from Sino Biological (HG11337-CY; Sino Biological, Beijing, China).

Techniques: Amplification

Journal: Cancers

Article Title: Enhancer of Zeste Homolog 2 Protects Mucosal Melanoma from Ferroptosis via the KLF14-SLC7A11 Signaling Pathway

doi: 10.3390/cancers16213660

Figure Lengend Snippet: Correlation of EZH2 gain to clinicopathologic features of mucosal melanoma.

Article Snippet: pCMV3-HA vector containing the human EZH2 coding sequence (HA- EZH2 ) was purchased from Sino Biological (HG11337-CY; Sino Biological, Beijing, China).

Techniques:

Journal: Cancers

Article Title: Enhancer of Zeste Homolog 2 Protects Mucosal Melanoma from Ferroptosis via the KLF14-SLC7A11 Signaling Pathway

doi: 10.3390/cancers16213660

Figure Lengend Snippet: Overall survival of melanoma patients in relation to EZH2 copy number variations. Comparison of the overall survival (OS) of tumors with different EZH2 copy number levels in melanoma subtypes was conducted by the Kaplan–Meier method. ( A ) all melanoma cases, n = 547. ( B ) acral melanoma cases, n = 252. ( C ) cutaneous melanoma cases, n = 147. ( D ) mucosal melanoma cases, n = 148. EZH2 -No gain was considered as samples with copy numbers less than or equal to 2.0. EZH2 gain was considered as samples with copy numbers greater than 2.0.

Article Snippet: pCMV3-HA vector containing the human EZH2 coding sequence (HA- EZH2 ) was purchased from Sino Biological (HG11337-CY; Sino Biological, Beijing, China).

Techniques: Comparison

Journal: Cancers

Article Title: Enhancer of Zeste Homolog 2 Protects Mucosal Melanoma from Ferroptosis via the KLF14-SLC7A11 Signaling Pathway

doi: 10.3390/cancers16213660

Figure Lengend Snippet: Univariate and multivariate analysis of EZH2 gain and clinicopathologic factors associated with overall survival in mucosal melanoma.

Article Snippet: pCMV3-HA vector containing the human EZH2 coding sequence (HA- EZH2 ) was purchased from Sino Biological (HG11337-CY; Sino Biological, Beijing, China).

Techniques: Mutagenesis

Journal: Cancers

Article Title: Enhancer of Zeste Homolog 2 Protects Mucosal Melanoma from Ferroptosis via the KLF14-SLC7A11 Signaling Pathway

doi: 10.3390/cancers16213660

Figure Lengend Snippet: Loss of EZH2 inhibits MM cell proliferation and progression in vitro and in vivo. ( A ) The expression of EZH2 was detected by RT-qPCR (top row) or Western blot assay (bottom row) after the knockdown of EZH2 . The data are presented as the mean ± SEM. n = 3, *** p < 0.001. ( B ) HMV-II, LM-MEL-53, and GAK cells with stable depletion of EZH2 or control were grown for 5 days, with cell numbers counted every day by CCK-8 assays. The changes in cell numbers were compared to day 0, and the mean ± SEM from 3 experiments was plotted. *** p < 0.001. ( C ) The proliferative abilities of stably EZH2 -depleted HMVII cells were measured with Ki-67 staining assay. Three experiments were conducted with mean ± SEM of percentage of Ki-67-positive cells plotted. Scale bar: 100 μm. *** p < 0.001. ( D ) The average sizes of xenograft tumors were measured every 3 days and plotted ( n = 5, error bars indicate mean ± SEM). * p < 0.05, *** p < 0.001.

Article Snippet: pCMV3-HA vector containing the human EZH2 coding sequence (HA- EZH2 ) was purchased from Sino Biological (HG11337-CY; Sino Biological, Beijing, China).

Techniques: In Vitro, In Vivo, Expressing, Quantitative RT-PCR, Western Blot, Knockdown, Control, CCK-8 Assay, Stable Transfection, Staining

Journal: Cancers

Article Title: Enhancer of Zeste Homolog 2 Protects Mucosal Melanoma from Ferroptosis via the KLF14-SLC7A11 Signaling Pathway

doi: 10.3390/cancers16213660

Figure Lengend Snippet: EZH2 knockdown enhances ferroptotic sensitivity in MM cells. ( A ) Cell viability response to treatment with apoptosis and ferroptosis inducers in cells with EZH2 knockdown. Cell viability was assessed after treatment with a range of concentrations of apoptosis inducer staurosporine (left panel), apoptosis inducer actinomycin D (middle panel), and ferroptosis inducer erastin (right panel). The mean ± SEM from 3 experiments was plotted. ( B ) Representative images of EZH2 knockdown effects on the viability of 3D spheroids formed by LM-MEL-53 cells in response to 4 µM erastin, as indicated by GFP fluorescence. Scale bars: 200 µm. ( C ) Bar graph showing viability of LM-MEL-53 cells with EZH2 knockdown, treated with 4 µM erastin or 4 µM erastin and 4 µM Ferrostatin-1. The data are presented as the mean ± SEM. n = 3, * p < 0.05, *** p < 0.001. ( D ) Bar graph demonstrating intracellular glutathione levels in EZH2 -depleted HMV-II and LM-MEL-53 cells. The data are presented as the mean ± SEM. n = 3, ** p < 0.01. ( E ) Lipid peroxidation was measured by flow cytometry after 5 μM CellROX Deep Red staining in EZH2 -depleted cells. The data are presented as the mean ± SEM. n = 3, *** p < 0.001. ( F ) The level of malondialdehyde in cells was determined by using a malondialdehyde kit after the knockdown of EZH2 . The data are presented as the mean ± SEM. n = 3, *** p < 0.001. ( G ) TEM was used to detect the mitochondrial morphology of ferroptotic cells. Scale bars: 1 μm (left column), and 500 nm (right column).

Article Snippet: pCMV3-HA vector containing the human EZH2 coding sequence (HA- EZH2 ) was purchased from Sino Biological (HG11337-CY; Sino Biological, Beijing, China).

Techniques: Knockdown, Fluorescence, Flow Cytometry, Staining

Journal: Cancers

Article Title: Enhancer of Zeste Homolog 2 Protects Mucosal Melanoma from Ferroptosis via the KLF14-SLC7A11 Signaling Pathway

doi: 10.3390/cancers16213660

Figure Lengend Snippet: Depletion of EZH2 stimulates ferroptosis through decreased SLC7A11 . ( A ) Volcano plot displaying differential expression from RNA-seq data. Red points indicate upregulated genes, blue points show downregulated genes, and grey points represent genes without significant changes. Vertical dashed lines mark fold change thresholds, and the horizontal line indicates the p -value cutoff for significance. ( B ) Venn diagram showing the significant overlap between RNA-seq data and FerrDb database. SCORE values were derived from FerrD. The SCORE values were listed below the Venn diagram, with SLC7A11 as the top hit ( C , D ). The expression of SLC7A11 was detected by RT-qPCR ( C ) or Western blot assay ( D ) with EZH2 knockdown. The data are presented as the mean ± SEM. n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001. ( E ) Representative phase-contrast images of EZH2 -depleted LM-MEL-53 cells, with or without SLC7A11 re-expression, treated with 4 μM erastin or 4 µM erastin and 4 µM Ferrostatin-1 ( n = 3). Scale bar: 200 μm.

Article Snippet: pCMV3-HA vector containing the human EZH2 coding sequence (HA- EZH2 ) was purchased from Sino Biological (HG11337-CY; Sino Biological, Beijing, China).

Techniques: Expressing, RNA Sequencing Assay, Derivative Assay, Quantitative RT-PCR, Western Blot, Knockdown

Journal: Cancers

Article Title: Enhancer of Zeste Homolog 2 Protects Mucosal Melanoma from Ferroptosis via the KLF14-SLC7A11 Signaling Pathway

doi: 10.3390/cancers16213660

Figure Lengend Snippet: EZH2 -mediated SLC7A11 upregulation is regulated by KLF14 . ( A ) Venn diagram of RNA-seq and ChIP-seq related genes showing that 73 genes were found to be potential target genes of EZH2 . ( B ) Heatmap showing the 6 transcription factors of the 73 potential target genes of EZH2 . ( C ) The binding of EZH2 to KLF14 promoter was detected after EZH2 overexpression by ChIP-qPCR. IgG as a negative control. The data are presented as the mean ± SEM. n = 3, ns, not significant, *** p < 0.001. ( D ) The protein level of KLF14 was detected with EZH2 depletion. ( E ) The protein level of SLC7A11 after transfection of KLF14 siRNA. ( F ) Schematic representation of the predicted KLF14 binding site within the SLC7A11 promoter. ( G ) The binding of KLF14 to SLC7A11 promoter was detected after KLF14 overexpression by ChIP-qPCR. IgG as a negative control. The data are presented as the mean ± SEM. n = 3, ns, not significant, *** p < 0.001. ( H ) Luciferase assay measuring SLC7A11 promoter activity before and after KLF14 binding site deletion in the absence or presence of KLF14 . Luciferase activities were normalized to Renilla luciferase activity. The data are presented as the mean ± SEM. n = 3, *** p < 0.001. ( I ) Western blot analysis of EZH2, KLF14, and SLC7A11 protein expression following dual knockdown of EZH2 and KLF14 . ( J ) The correlations between EZH2 and SLC7A11 protein expression in MM patients were analyzed by Pearson correlation analysis ( n = 55). Representative images from immunohistochemical staining of EZH2 and SLC7A11 protein expression. The staining score for each sample, counting the intensity of the staining, was graded as 0, 1, 2, and 3 (“0” as negative, and “3” as the strongest). Scale bar: 100 μm.

Article Snippet: pCMV3-HA vector containing the human EZH2 coding sequence (HA- EZH2 ) was purchased from Sino Biological (HG11337-CY; Sino Biological, Beijing, China).

Techniques: RNA Sequencing Assay, ChIP-sequencing, Binding Assay, Over Expression, Negative Control, Transfection, Luciferase, Activity Assay, Western Blot, Expressing, Knockdown, Immunohistochemical staining, Staining

Journal: Cancers

Article Title: Enhancer of Zeste Homolog 2 Protects Mucosal Melanoma from Ferroptosis via the KLF14-SLC7A11 Signaling Pathway

doi: 10.3390/cancers16213660

Figure Lengend Snippet: Effects of combined EZH2 inhibitor and ferroptosis inducer treatment in MM. ( A – C ) Evaluation of combinatorial treatment effects on melanoma organoid models. ( A ) Schematic representation of the experimental design for combinatorial treatment with MS8815 and erastin on melanoma organoid models. ( B ) Representative images of melanoma organoids treated with DMSO (vehicle control), MS8815, erastin, and a combination of MS8815 and erastin, showing morphological changes ( n = 3). Scale bar: 100 µm. ( C ) Quantification of cellular ATP levels as a measure of cell viability post-treatment (*** p < 0.001 compared to vehicle control). ( D – F ) Effects of combined MS8815 and erastin on tumor growth in a MM PDX model. ( D ) Tumor volume was measured over an 18-day period. ( E ) Tumors were weighed and plotted ( n = 5). Data were expressed as mean ± SEM. * p < 0.05, *** p < 0.001. ( F ) Representative images from H&E and immunohistochemical staining of Ki-67 protein expression. Scale bars: 100 μm.

Article Snippet: pCMV3-HA vector containing the human EZH2 coding sequence (HA- EZH2 ) was purchased from Sino Biological (HG11337-CY; Sino Biological, Beijing, China).

Techniques: Control, Immunohistochemical staining, Staining, Expressing