Journal: Frontiers in Genetics

Article Title: Chrysin Modulates Aberrant Epigenetic Variations and Hampers Migratory Behavior of Human Cervical (HeLa) Cells

doi: 10.3389/fgene.2021.768130

Figure Lengend Snippet: Relative expression of TSGs and genes related to migration and metastasis. The values are taken as mean of three experiments ±SD ( p ≤ 0.05).

Article Snippet: MTA2 , Hs00191018_m1 , metastasis associated 2 , 0.28 , 0.14.

Techniques: Expressing, Migration

Journal: Cancer Science

Article Title: MTA2 promotes the metastasis of esophageal squamous cell carcinoma via EIF4E‐Twist feedback loop

doi: 10.1111/cas.14778

Figure Lengend Snippet: MTA2 overexpression in ESCC tissues correlates with ESCC aggressiveness. A, Expression of MTA2 was analyzed by qRT‐PCR in ESCC tissues (n = 98), adjacent non‐tumor tissues (n = 98), and normal esophageal epithelial tissues (n = 34). B‐D, The correlation between MTA2 expression and primary tumor invasion depth (B), TNM stage (C), lymph node metastasis (D). E, Expression of MTA2 was analyzed by IHC in primary tumor tissues and the corresponding lymph node metastatic tumors tissues from 3 patients. F, Effect of the MTA2 expression level on clinical prognosis was analyzed by Kaplan‐Meier survival analysis. * P < .05, ** P < .01, *** P < .001

Article Snippet: The cells were then incubated with antibodies, including MTA2, E‐cadherin, N‐cadherin, and vimentin (CST) for 2 h at room temperature, followed by further incubation at room temperature for 1 h with rabbit IgG (Alexa Fluor 546, green).

Techniques: Over Expression, Expressing, Quantitative RT-PCR

Journal: Cancer Science

Article Title: MTA2 promotes the metastasis of esophageal squamous cell carcinoma via EIF4E‐Twist feedback loop

doi: 10.1111/cas.14778

Figure Lengend Snippet: Correlation between MTA2 expression and clinicopathological features in 98 patients with ESCC

Article Snippet: The cells were then incubated with antibodies, including MTA2, E‐cadherin, N‐cadherin, and vimentin (CST) for 2 h at room temperature, followed by further incubation at room temperature for 1 h with rabbit IgG (Alexa Fluor 546, green).

Techniques: Expressing

Journal: Cancer Science

Article Title: MTA2 promotes the metastasis of esophageal squamous cell carcinoma via EIF4E‐Twist feedback loop

doi: 10.1111/cas.14778

Figure Lengend Snippet: MTA2 is required for ESCC cell proliferation, migration and invasion in vitro. A, Wound healing assay were performed in KYSE30 and KYSE510 cells transfected with siRNA. B and C, Transwell and Matrigel assays were performed in control and MTA2‐depleted KYSE30 and KYSE510 cells. D, Wound healing assays were performed in KYSE30 and KYSE510 cells transfected with different vectors. E and F, Transwell and Matrigel assays were performed in control and MTA2‐overexpressed KYSE30 and KYSE510 cells. * P < .05, ** P < .01

Article Snippet: The cells were then incubated with antibodies, including MTA2, E‐cadherin, N‐cadherin, and vimentin (CST) for 2 h at room temperature, followed by further incubation at room temperature for 1 h with rabbit IgG (Alexa Fluor 546, green).

Techniques: Migration, In Vitro, Wound Healing Assay, Transfection, Control

Journal: Cancer Science

Article Title: MTA2 promotes the metastasis of esophageal squamous cell carcinoma via EIF4E‐Twist feedback loop

doi: 10.1111/cas.14778

Figure Lengend Snippet: MTA2 promotes the growth and metastasis of transplanted tumors in vivo. A, C, KYSE30 cells stably downregulated (A) or overexpressed (C) MTA2 were subcutaneously injected into BALB/c nude mice. Representative images of mice from different treatment groups 6 wk after injections are shown. Tumor growth and tumor weight were analyzed. B and D, Representative immunohistochemical images showing the intensity of Ki‐67 and CD31 expression (brown) in xenografted tumors from different groups. E, KYSE30 cells stably downregulated MTA2 were slowly injected into BALB/c nude mice via the tail vein. Representative images of HE staining of lung metastasis. ** P < .01, *** P < .001

Article Snippet: The cells were then incubated with antibodies, including MTA2, E‐cadherin, N‐cadherin, and vimentin (CST) for 2 h at room temperature, followed by further incubation at room temperature for 1 h with rabbit IgG (Alexa Fluor 546, green).

Techniques: In Vivo, Stable Transfection, Injection, Immunohistochemical staining, Expressing, Staining

Journal: Cancer Science

Article Title: MTA2 promotes the metastasis of esophageal squamous cell carcinoma via EIF4E‐Twist feedback loop

doi: 10.1111/cas.14778

Figure Lengend Snippet: MTA2 is required for epithelial‐mesenchymal transition in ESCC cells. A and B, Western blot analysis of the expression of EMT markers in MTA2‐depleted or MTA2‐overexpressed KYSE30 and KYSE510 cells. C, Immunofluorescence staining for EMT markers in KYSE30 cells. transfected with siRNAs. D, Expression of epithelial marker E‐cadherin and the mesenchymal markers N‐cadherin and vimentin in mice tumor tissues was examined by IHC

Article Snippet: The cells were then incubated with antibodies, including MTA2, E‐cadherin, N‐cadherin, and vimentin (CST) for 2 h at room temperature, followed by further incubation at room temperature for 1 h with rabbit IgG (Alexa Fluor 546, green).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Transfection, Marker

Journal: Cancer Science

Article Title: MTA2 promotes the metastasis of esophageal squamous cell carcinoma via EIF4E‐Twist feedback loop

doi: 10.1111/cas.14778

Figure Lengend Snippet: EIF4E is a function target of MTA2. A, Heatmap representation of the fold change in gene expression as determined by transcriptome analysis in MTA2‐silenced cells and control cells (KYSE30). B, KEGG pathway analysis of MTA2‐regulated genes. Pathways showing enrichment are presented. C, Expression of cancer‐related genes after MTA2 knockdown was detected by qRT‐PCR. D, Expression of EIF4E was analyzed by qRT‐PCR in esophageal squamous cell carcinoma tissues (n = 60) and adjacent non‐tumor tissues (n = 60). E‐G, The correlation between EIF4E expression and primary tumor invasion depth (E), lymph node metastasis (F), and TNM stage (G). H, Effect of EIF4E expression level on clinical prognosis was analyzed by Kaplan‐Meier survival analysis. * P < .05, ** P < .01, *** P < .001

Article Snippet: The cells were then incubated with antibodies, including MTA2, E‐cadherin, N‐cadherin, and vimentin (CST) for 2 h at room temperature, followed by further incubation at room temperature for 1 h with rabbit IgG (Alexa Fluor 546, green).

Techniques: Gene Expression, Control, Expressing, Knockdown, Quantitative RT-PCR

Journal: Cancer Science

Article Title: MTA2 promotes the metastasis of esophageal squamous cell carcinoma via EIF4E‐Twist feedback loop

doi: 10.1111/cas.14778

Figure Lengend Snippet: MTA2‐mediated promotion of malignancy in ESCC is mediated by EIF4E. A, Western blot assay was performed to examine the expression of EIF4E after MTA2 knockdown or overexpressed in KYSE30 and KYSE510 cells. B, Expression of EIF4E in mice tumor tissues from different treat groups was examined by IHC. C, Correlation between MTA2 and EIF4E expression in ESCC tissues was analyzed by Spearman test. D, Transwell assays were performed in control and EIF4E‐knockdown KYSE30 and KYSE510 cells. The numbers of cells that migrated or invaded were counted in 5 different fields. E, Transwell assays were performed in control and EIF4E‐overexpressed KYSE30 and KYSE510 cells. The numbers of cells that migrated or invaded were counted in 5 different fields. F, Rescue experiments with transwell assays were performed in KYSE30 cells co‐transfected with shMTA2 and EIF4E or MTA2 and si‐EIF4E#1. The numbers of cells that migrated or invaded were counted in 5 different fields. ** P < .01, *** P < .001

Article Snippet: The cells were then incubated with antibodies, including MTA2, E‐cadherin, N‐cadherin, and vimentin (CST) for 2 h at room temperature, followed by further incubation at room temperature for 1 h with rabbit IgG (Alexa Fluor 546, green).

Techniques: Western Blot, Expressing, Knockdown, Control, Transfection

Journal: Cancer Science

Article Title: MTA2 promotes the metastasis of esophageal squamous cell carcinoma via EIF4E‐Twist feedback loop

doi: 10.1111/cas.14778

Figure Lengend Snippet: MTA2 promotes epithelial‐mesenchymal transition in ESCC via regulation of EIF4E. A, Western blot analysis of the expression of EMT markers in EIF4E‐overexpressed KYSE30 and KYSE510 cells. B, Western blot analysis of the expression of EMT markers in EIF4E‐depleted KYSE30 and KYSE510 cells. C, Overexpression of EIF4E reversed the effect of MTA2 downregulation on EMT transformation. D, Knocking down EIF4E expression reversed the effect of MTA2 upregulation on EMT transformation

Article Snippet: The cells were then incubated with antibodies, including MTA2, E‐cadherin, N‐cadherin, and vimentin (CST) for 2 h at room temperature, followed by further incubation at room temperature for 1 h with rabbit IgG (Alexa Fluor 546, green).

Techniques: Western Blot, Expressing, Over Expression, Transformation Assay

Journal: Cancer Science

Article Title: MTA2 promotes the metastasis of esophageal squamous cell carcinoma via EIF4E‐Twist feedback loop

doi: 10.1111/cas.14778

Figure Lengend Snippet: MTA2 and EIF4E‐Twist formed a positive feedback to repress E‐cadherin expression. A, Transwell assays were performed in control and Twist‐overexpressed KYSE30 and KYSE510 cells. The numbers of cells that migrated or invaded were counted in 5 different fields. B, Western blot analysis of the expression of EMT markers in Twist‐overexpressed KYSE30 and KYSE510 cells. C, Coimmunoprecipitation assays were performed in control and MTA2‐overexpressed KYSE30 cells. IgG was used as a negative control. D, Endogenous MTA2 and Twist were coimmunoprecipitated from KYSE30 cells and the bound endogenous MTA2 and Twist were examined by immunoblotting. E, Twist recognized consensus site was identified in the promoter region of E‐cadherin using a bioinformatics website ( http://jaspar.genereg.net/ ). The number represents the nucleotide position relative to the transcription start site (+1). F, ChIP Re‐ChIP assays were performed in KYSE30 cells to examine the recruitment of Twist and MTA2 onto E‐cadherin promoter. G, ChIP assays were performed in KYSE30 cells to examine the promoter acetylation level of E‐cadherin with or without MTA2. Ac, acetylated lysine. H, Coimmunoprecipitation assays were performed to detect the acetylation of E‐cadherin in MTA2 overexpression or knockdown KYSE30 cells. Ac, acetylated lysine. I, Effect of EIF4E on the expression of Twist was analyzed by western blot. J, The influence of MTA2 and EIF4E on the expression of Twist was examined by western blot. K, Rescue experiment with western blot analysis on the influence of MTA2 and Twist on the expression of EMT markers in KYSE30 cells. L, Schematic model of MTA2 interaction with EIF4E and Twist involved in pro‐metastatic function. * P < .05, ** P < .01

Article Snippet: The cells were then incubated with antibodies, including MTA2, E‐cadherin, N‐cadherin, and vimentin (CST) for 2 h at room temperature, followed by further incubation at room temperature for 1 h with rabbit IgG (Alexa Fluor 546, green).

Techniques: Expressing, Control, Western Blot, Negative Control, Over Expression, Knockdown

Journal: Cellular and molecular biology (Noisy-le-Grand, France)

Article Title: lncRNA PCAT1 might coordinate ZNF217 to promote CRC adhesion and invasion through regulating MTA2/MTA3/Snai1/E-cadherin signaling.

doi: 10.14715/cmb/2021.67.4.1

Figure Lengend Snippet: Figure 5. The protein expressions of MTA2, MTA3, SNAI1 and E-cadherin in CRC tissue and cell lines. (a) The upregulation of MTA2, MTA3, and SNAI1, and downregulation of E-cadherin in stage 0-I and stage II-III CRC tissues samples. (b) The upregulation of MTA2, MTA3, and SNAI1, and downregulation of E-cadherin in both HT29 and HCT8 cells. * p < 0 05, * p < 0 01 and ***p < 0 001 vs paracancerous CRC or FHC cell line. # p < 0 01, ## p < 0 01, and ### p < 0 001 vs stage 0-I CRC or HT29 cell line.

Article Snippet: Subsequently, the membranes were incubated overnight at 4 °C with Rabbit polyclonal MTA2 (1:3000, 17554-1-AP, Proteintech), MTA3 (1:1000, 14682-1-AP, Proteintech), SNAI1 (1:5000, 20874-1- AP, Proteintech), and mouse anti-human E-cadherin (1:2000, 60335-1-Ig, Proteintech) and beta-actin as internal reference (1:8000, 60008-1-Ig, Proteintech).

Techniques:

Journal: Cellular and molecular biology (Noisy-le-Grand, France)

Article Title: lncRNA PCAT1 might coordinate ZNF217 to promote CRC adhesion and invasion through regulating MTA2/MTA3/Snai1/E-cadherin signaling.

doi: 10.14715/cmb/2021.67.4.1

Figure Lengend Snippet: Figure 6. ZNF217 knockdown reduces MTA2, MTA3, and E-cadherin proteins expressions, and increases SNAI1 expression in both HT29 (a) and HCT8 (b) cells. * p < 0 05, * p < 0 01 and ***p < 0 001 vs si-NC.

Article Snippet: Subsequently, the membranes were incubated overnight at 4 °C with Rabbit polyclonal MTA2 (1:3000, 17554-1-AP, Proteintech), MTA3 (1:1000, 14682-1-AP, Proteintech), SNAI1 (1:5000, 20874-1- AP, Proteintech), and mouse anti-human E-cadherin (1:2000, 60335-1-Ig, Proteintech) and beta-actin as internal reference (1:8000, 60008-1-Ig, Proteintech).

Techniques: Knockdown, Expressing

Journal: The Journal of Biological Chemistry

Article Title: Transcription suppression is mediated by the HDAC1–Sin3 complex in Xenopus nucleoplasmic extract

doi: 10.1016/j.jbc.2022.102578

Figure Lengend Snippet: The Sin3 deacetylase complex promotes transcription suppression. A , sequential IP schematic. B , mock, HDAC1, or HDAC2 IPs were performed in NPE. The supernatants from HDAC1 or HDAC2 IPs were then used for a second round of IPs using the converse antibody. Isolated proteins were then analyzed by Western blot with the indicated antibodies (n = 3). 10% of total reaction sample (IN), supernatant (S), pellet (P). C , NPE was immunodepleted using preimmune (ΔMock) or MTA2 (ΔMTA2) antibodies. Depleted extracts were analyzed by Western blot using the indicated antibodies. D , pActin was incubated in mock- or MTA2-depleted extract. RNA was isolated and quantified after 120 min (n = 2). E , NPE was immunodepleted using preimmune (ΔMock) or Sin3a (ΔSin3a) antibodies. Depleted extracts were analyzed by Western blot using the indicated antibodies. F , pActin was incubated in mock- or Sin3a-depleted extract. RNA was isolated and quantified after 120 min (n = 2). G , NPE was immunodepleted using preimmune (ΔMock) or HDAC1 (ΔHDAC1) antibodies. HDAC1-depleted extract was then supplemented with immunoprecipitated proteins recovered by preimmune (+Mock IP) or Sin3a (+Sin3a IP) IP. pActin was incubated in each extract and RNA was isolated and quantified after 120 min (n = 2). Student t test: p -value < 0.05 (∗), p -value < 0.01 (∗∗), p -value < 0.001 (∗∗∗), not significant (n.s.). Error bars represent ±1 SD. IP, immunoprecipitation; NPE, nucleoplasmic extract.

Article Snippet: Commercial antibodies were used to detect Histone H3 (ThermoFisher PA5-16183), H4K5ac (Abclonal A15233), H4K8ac (Abclonal A7258), H4K16ac (Abclonal A5280), H3K27ac (Abclonal A7253), MTA2 (Novus Biologicals NB100-56483SS), CoREST (Millipore 07-455), H2AK5ac (Thermofisher 720070), H2BK20ac (Millipore Sigma 07-347), H3K9/14ac (ThermoFisher 49-1010), H3K23ac (Cell Signaling 8848), and RNAPII (Bethyl Laboratories A300-653A).

Techniques: Histone Deacetylase Assay, Isolation, Western Blot, Incubation, Immunoprecipitation