Journal: Journal of Immunology Research

Article Title: HLA-A2-Restricted Epitopes Identified from MTA1 Could Elicit Antigen-Specific Cytotoxic T Lymphocyte Response

doi: 10.1155/2018/2942679

Figure Lengend Snippet: Protein expression of MTA1 by Western Blot analysis. The protein expression of MTA1 (81 kDa) was evaluated in four human cancer cell lines. β -Actin (42 kDa) was used as loading control, and rabbit polyclonal anti-MTA1 antibody (cat: bs-1412R, dilutions: 1 : 1500, Bioss Antibodies, China) or rabbit polyclonal anti- β -actin antibody (cat: bs-0061R, dilutions: 1 : 50,000, Bioss Antibodies, China) was used in this assay. (a) The bands representing MTA1 and β -actin in each cancer cell line. (b) Gray analysis of the MTA1 protein band intensities (relative to the β -actin band intensity) was carried out by Image J.

Article Snippet: Further, the membranes were incubated overnight at 4°C with the indicated primary antibody—rabbit anti-MTA1 antibody (cat: bs-1412R, dilutions: 1 : 1500, Bioss Antibodies, China/cat: 5646, dilutions: 1 : 500, Cell Signaling Technology, USA) or rabbit polyclonal anti- β -actin antibody (cat: bs-0061R, dilutions: 1 : 50,000, Bioss Antibodies, China)—and then washed three times in TBST.

Techniques: Expressing, Western Blot

Journal: Journal of Immunology Research

Article Title: HLA-A2-Restricted Epitopes Identified from MTA1 Could Elicit Antigen-Specific Cytotoxic T Lymphocyte Response

doi: 10.1155/2018/2942679

Figure Lengend Snippet: IFN- γ release and lyse target cell ability of peptide-induced CTLs. PBMCs from six HLA-A ∗ 02 + healthy donors were collected and induced by the indicated peptides (10 μ g/ml) three times in RPMI 1640 medium supplemented with 3 μ g/ml β 2-M (once at each stimulation), interleukin-2 (50 U/ml, on day 3 and 1 day after each stimulation), and 10% FCS for 21 days. CTLs induced by COX-2 321–329 were used as positive controls. On day 21, the CTLs were collected, and the IFN- γ secretion and cytotoxic activity of the CTLs were measured. (a) CTL secretion of IFN- γ was detected by the ELISPOT assay ( n = 6). T2 cells loaded with/without peptide (50 μ g/ml) for 4 h were used as the stimulating cells. LDH cytotoxicity assays were performed ( n = 3) using the following target cell lines: (b) SW620 (HLA-A2 + , MTA1 + ), (c) MDA-MB-231 (HLA-A2 + , MTA1 + ), (d) SW620 incubated with an anti-human HLA-A2 antibody (HLA-A2 − , MTA1 + ), (e) HT-29 (HLA-A2 − , MTA1 + ), (f) MCF-7 (HLA-A2 + , MTA1 + ), (g) HUVEC (HLA-A2 + , MTA1 low ), and (h) Het-1A (HLA-A2 + , MTA1 low ). CTLs induced by PBS were used as the negative control. Data represent means ± standard deviation (SD), ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 vs. the PBS group.

Article Snippet: Further, the membranes were incubated overnight at 4°C with the indicated primary antibody—rabbit anti-MTA1 antibody (cat: bs-1412R, dilutions: 1 : 1500, Bioss Antibodies, China/cat: 5646, dilutions: 1 : 500, Cell Signaling Technology, USA) or rabbit polyclonal anti- β -actin antibody (cat: bs-0061R, dilutions: 1 : 50,000, Bioss Antibodies, China)—and then washed three times in TBST.

Techniques: Activity Assay, Enzyme-linked Immunospot, Incubation, Negative Control, Standard Deviation

Journal: Journal of Immunology Research

Article Title: HLA-A2-Restricted Epitopes Identified from MTA1 Could Elicit Antigen-Specific Cytotoxic T Lymphocyte Response

doi: 10.1155/2018/2942679

Figure Lengend Snippet: Specific lysis of target cells and secretion of IFN- γ by induced CTLs in immunizing HLA-A2.1/K b transgenic mice immunized with the indicated peptides. Peptide (100 μ g) and the IA b -restricted HBV-core 128 T helper epitope (140 μ g) emulsified in IFA were used to immunize HLA-A2.1/K b transgenic mice on days 0, 5, and 10. PBS only or T helper peptide emulsified in IFA was used as the negative control. On day 11, we isolated splenocytes from the mice and restimulated the cells with 10 μ g/ml peptide and 10 U/ml rmIL-2 to repriming these splenocytes in vitro for additional 6 days. An LDH assay was performed to assess the lysis activity of the induced CTLs toward (a) SW620 cells (HLA-A2 + , MTA1 + ), (b) T2 cells pulsed with each peptide, and (c) SW620 cells incubated with an anti-human HLA-A2 antibody (HLA-A2 − , MTA1 + ). IFN- γ secretion was detected by the ELISPOT assay. (d) T2 cells incubated with each peptide and an irrelevant peptide (50 μ g/ml) for 4 h were used as the stimulating cells. Each sample was measured in triplicate. Data represent means ± SD ( n = 5). ∗ P < 0.05, ∗∗ P < 0.05, and ∗∗∗ P < 0.001 vs. the T helper epitope group.

Article Snippet: Further, the membranes were incubated overnight at 4°C with the indicated primary antibody—rabbit anti-MTA1 antibody (cat: bs-1412R, dilutions: 1 : 1500, Bioss Antibodies, China/cat: 5646, dilutions: 1 : 500, Cell Signaling Technology, USA) or rabbit polyclonal anti- β -actin antibody (cat: bs-0061R, dilutions: 1 : 50,000, Bioss Antibodies, China)—and then washed three times in TBST.

Techniques: Lysis, Transgenic Assay, Negative Control, Isolation, In Vitro, Lactate Dehydrogenase Assay, Activity Assay, Incubation, Enzyme-linked Immunospot

Journal: Journal of Immunology Research

Article Title: HLA-A2-Restricted Epitopes Identified from MTA1 Could Elicit Antigen-Specific Cytotoxic T Lymphocyte Response

doi: 10.1155/2018/2942679

Figure Lengend Snippet: Specific T cells induced by MTA1 (1–283) secrete IFN- γ and lyse target cells. PBMCs from healthy HLA-A ∗ 02 + donors were isolated and stimulated with protein MTA1 (1–283) (at final concentrations of 0, 1, 5, 10, 30, and 50 μ g/ml) in RPMI 1640 medium supplemented with 50 U/ml interleukin-2 and 10% FCS once a week for 21 days. On day 21, the induced T cells were collected; (a) IFN- γ secretion was assessed by the ELISPOT assay; ∗ P < 0.05 and ∗∗∗ P < 0.001 between two groups. (b) Cytotoxic activity was assessed by the LDH assay using SW620 cells (HLA-A2 + , MTA1 + ) as the target cells. T cells induced by PBS were used as negative controls. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 vs. the control group. Data represent means ± SD ( n = 4).

Article Snippet: Further, the membranes were incubated overnight at 4°C with the indicated primary antibody—rabbit anti-MTA1 antibody (cat: bs-1412R, dilutions: 1 : 1500, Bioss Antibodies, China/cat: 5646, dilutions: 1 : 500, Cell Signaling Technology, USA) or rabbit polyclonal anti- β -actin antibody (cat: bs-0061R, dilutions: 1 : 50,000, Bioss Antibodies, China)—and then washed three times in TBST.

Techniques: Isolation, Enzyme-linked Immunospot, Activity Assay, Lactate Dehydrogenase Assay

Journal: Journal of Immunology Research

Article Title: HLA-A2-Restricted Epitopes Identified from MTA1 Could Elicit Antigen-Specific Cytotoxic T Lymphocyte Response

doi: 10.1155/2018/2942679

Figure Lengend Snippet: Specific lysis of various cell lines and IFN- γ secretion by induced T cells in HLA-A2.1/K b transgenic mice immunized with MTA1 (1–283) . MTA1 (1–283) (100 μ g/ml) or the same volume of PBS (negative control) emulsified in IFA was used to immunize HLA-A2.1/K b transgenic mice on days 0 and 7. On day 14, the mice were sacrificed, and their splenocytes were collected and restimulated for another 6 days with the peptide pool (10 μ g/ml) and 50 U/ml rmIL-2 in vitro . (a) Serum was collected from the immunized mice on day 14, and the serum IFN- γ concentration was measured by ELISA. (b) The percentage of IFN- γ secreting peptide-specific CD8 + T cells induced by MTA1 (1–283) and PBS was analyzed by an intracellular staining assay. (c) The percentages of IFN-γ + CD4 + T cells in the MTA1 (1–283) and PBS groups were determined. (d) Specific lysis of target cells SW620 cells (HLA-A2 + , MTA1 + ) was assessed by the LDH cytotoxicity assay. ∗ P < 0.05 and ∗∗∗ P < 0.001. P < 0.05 vs. the control group. Data represent means ± SD ( n = 5).

Article Snippet: Further, the membranes were incubated overnight at 4°C with the indicated primary antibody—rabbit anti-MTA1 antibody (cat: bs-1412R, dilutions: 1 : 1500, Bioss Antibodies, China/cat: 5646, dilutions: 1 : 500, Cell Signaling Technology, USA) or rabbit polyclonal anti- β -actin antibody (cat: bs-0061R, dilutions: 1 : 50,000, Bioss Antibodies, China)—and then washed three times in TBST.

Techniques: Lysis, Transgenic Assay, Negative Control, In Vitro, Concentration Assay, Enzyme-linked Immunosorbent Assay, Staining, LDH Cytotoxicity Assay

Journal: Mediators of Inflammation

Article Title: Ellagic Acid Alleviates Rheumatoid Arthritis in Rats through Inhibiting MTA1/HDAC1-Mediated Nur77 Deacetylation

doi: 10.1155/2021/6359652

Figure Lengend Snippet: EA reduced MTA1 expression in MH7A cells. (a) The molecular formula of EA. (b) MH7A cells were incubated with different concentrations (0, 10, 25, 50, and 100 μ M) of EA. MH7A cell viability was measured by using CCK-8 assay. ∗ P < 0.05 compared with 0 μ M EA group. (c) MH7A cells were treated with 10 ng/mL TNF- α to induce inflammation injury and then incubated with different concentrations of EA. Western blot analysis was performed to evaluate the expression of MTA1 protein. Data from at least three replicate experiments were presented as mean ± SD. N = 5. ∗∗ P < 0.01 compared with the first group, # P < 0.05, ## P < 0.01 compared with the second group.

Article Snippet: Then, equal amount of protein was separated on 10% SDS-PAGE, and the protein bands were transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, MA, USA).The membranes were blocked with 5% nonfat milk for 2 h at room temperature and then incubated overnight at 4°C with the following primary antibodies: rabbit polyclonal anti-MTA1 antibody (1 : 1000, ab71153, Abcam), rabbit polyclonal anti-HDAC1 antibody (1 : 1000, ab19845, Abcam), rabbit polyclonal anti-Nur77 antibody (1 : 1000, ab13851, Abcam), and rabbit polyclonal anti-GAPDH antibody (1 : 2500, ab9485, Abcam).

Techniques: Expressing, Incubation, CCK-8 Assay, Western Blot

Journal: Mediators of Inflammation

Article Title: Ellagic Acid Alleviates Rheumatoid Arthritis in Rats through Inhibiting MTA1/HDAC1-Mediated Nur77 Deacetylation

doi: 10.1155/2021/6359652

Figure Lengend Snippet: EA inhibited proliferation, inflammation, and oxidative stress and promoted apoptosis by downregulating MTA1 in MH7A cells. TNF- α -treated MH7A cells were treated with 50 μ M EA alone or together with pcDNA-MTA1. (a, b) The expression of MTA1 protein was detected by Western blot analysis. (c) CCK-8 assay was performed to measure MH7A cell proliferation. (d, e) MH7A cell apoptosis was measured by flow cytometry. (f, g) The levels of IL-6 and IL-1 β in cell culture supernatant were detected by ELISA. (h, i) The levels of SOD and MDA in MH7A cells were examined by commercial kits. Data from at least three replicate experiments were presented as mean ± SD. N = 5. ∗∗ P < 0.01.

Article Snippet: Then, equal amount of protein was separated on 10% SDS-PAGE, and the protein bands were transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, MA, USA).The membranes were blocked with 5% nonfat milk for 2 h at room temperature and then incubated overnight at 4°C with the following primary antibodies: rabbit polyclonal anti-MTA1 antibody (1 : 1000, ab71153, Abcam), rabbit polyclonal anti-HDAC1 antibody (1 : 1000, ab19845, Abcam), rabbit polyclonal anti-Nur77 antibody (1 : 1000, ab13851, Abcam), and rabbit polyclonal anti-GAPDH antibody (1 : 2500, ab9485, Abcam).

Techniques: Expressing, Western Blot, CCK-8 Assay, Flow Cytometry, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Mediators of Inflammation

Article Title: Ellagic Acid Alleviates Rheumatoid Arthritis in Rats through Inhibiting MTA1/HDAC1-Mediated Nur77 Deacetylation

doi: 10.1155/2021/6359652

Figure Lengend Snippet: MTA1 interacted with HDAC1 and promoted HDAC1 expression. (a) The interaction between MTA1 protein and HDAC1 protein was verified by Co-IP assay. (b–d) pcDNA-MTA1, si-MTA1, and their negative controls were transfected into MH7A cells, respectively. The expression of MTA1 and HDAC1 was measured by Western blot analysis. (e, f) TNF- α -treated MH7A cells were treated with 50 μ M EA alone or together with pcDNA-MTA1. The expression of HDAC1 was measured by Western blot analysis. Data from at least three replicate experiments were presented as mean ± SD. N = 5. ∗∗ P < 0.01.

Article Snippet: Then, equal amount of protein was separated on 10% SDS-PAGE, and the protein bands were transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, MA, USA).The membranes were blocked with 5% nonfat milk for 2 h at room temperature and then incubated overnight at 4°C with the following primary antibodies: rabbit polyclonal anti-MTA1 antibody (1 : 1000, ab71153, Abcam), rabbit polyclonal anti-HDAC1 antibody (1 : 1000, ab19845, Abcam), rabbit polyclonal anti-Nur77 antibody (1 : 1000, ab13851, Abcam), and rabbit polyclonal anti-GAPDH antibody (1 : 2500, ab9485, Abcam).

Techniques: Expressing, Co-Immunoprecipitation Assay, Transfection, Western Blot

Journal: Mediators of Inflammation

Article Title: Ellagic Acid Alleviates Rheumatoid Arthritis in Rats through Inhibiting MTA1/HDAC1-Mediated Nur77 Deacetylation

doi: 10.1155/2021/6359652

Figure Lengend Snippet: MTA1 knockdown inhibited proliferation, inflammation, and oxidative stress and promoted apoptosis by downregulating HDAC1 in MH7A cells. TNF- α -treated MH7A cells were transfected with si-MTA1 alone or together with pcDNA-HDAC1. (a–c) The protein expression of MTA1 and HDAC1 was detected by Western blot analysis. (d) MH7A cell proliferation was measured by CCK-8 assay. (e, f) Flow cytometry was performed to measure MH7A cell apoptosis. (g, h) The levels of IL-6 and IL-1 β in cell culture supernatant were detected by ELISA. (i, j) The levels of SOD and MDA in MH7A cells were examined by commercial kits. Data from at least three replicate experiments were presented as mean ± SD. N = 5. ∗∗ P < 0.01.

Article Snippet: Then, equal amount of protein was separated on 10% SDS-PAGE, and the protein bands were transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, MA, USA).The membranes were blocked with 5% nonfat milk for 2 h at room temperature and then incubated overnight at 4°C with the following primary antibodies: rabbit polyclonal anti-MTA1 antibody (1 : 1000, ab71153, Abcam), rabbit polyclonal anti-HDAC1 antibody (1 : 1000, ab19845, Abcam), rabbit polyclonal anti-Nur77 antibody (1 : 1000, ab13851, Abcam), and rabbit polyclonal anti-GAPDH antibody (1 : 2500, ab9485, Abcam).

Techniques: Knockdown, Transfection, Expressing, Western Blot, CCK-8 Assay, Flow Cytometry, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Mediators of Inflammation

Article Title: Ellagic Acid Alleviates Rheumatoid Arthritis in Rats through Inhibiting MTA1/HDAC1-Mediated Nur77 Deacetylation

doi: 10.1155/2021/6359652

Figure Lengend Snippet: EA alleviated RA progression in CIA rats. Wistar rats were randomly divided into five groups ( n = 8 per group): control, CIA, CIA + EA (25 mg/kg), CIA + EA (50 mg/kg), and CIA + EA (100 mg/kg). The rats were immunized by intradermally injecting the collagen emulsion to establish CIA rat model, and EA was orally administered. (a–d) The expression of MTA1, HDAC1, and Nur77 in synovial tissues of rats was measured by using Western blot analysis. (e) The arthritis index of rats in each group was detected once a week. (f) The paw swelling of rats in each group was detected once a week. (g) Effects of EA on histopathological changes in the synovial tissues of CIA rats, using HE staining. (h–j) The levels of TNF- α , IL-1 β , and IL-6 in synovial tissues were examined by using ELISA. Data from at least three replicate experiments were presented as mean ± SD. ∗∗ P < 0.01 compared with the control group; # P < 0.05 and ## P < 0.01, compared with the CIA group.

Article Snippet: Then, equal amount of protein was separated on 10% SDS-PAGE, and the protein bands were transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, MA, USA).The membranes were blocked with 5% nonfat milk for 2 h at room temperature and then incubated overnight at 4°C with the following primary antibodies: rabbit polyclonal anti-MTA1 antibody (1 : 1000, ab71153, Abcam), rabbit polyclonal anti-HDAC1 antibody (1 : 1000, ab19845, Abcam), rabbit polyclonal anti-Nur77 antibody (1 : 1000, ab13851, Abcam), and rabbit polyclonal anti-GAPDH antibody (1 : 2500, ab9485, Abcam).

Techniques: Control, Emulsion, Expressing, Western Blot, Staining, Enzyme-linked Immunosorbent Assay

Journal: Theranostics

Article Title: LncRNA-MTA2TR functions as a promoter in pancreatic cancer via driving deacetylation-dependent accumulation of HIF-1α

doi: 10.7150/thno.34559

Figure Lengend Snippet: MTA2TR facilitates the in vivo invasion and proliferation of BxPC-3 cells. (A-F) BxPC-3 cells transfected using lentiviruses encoding siMTA2TR#1 (LV-siMTA2TR#1), siMTA2TR#2 (LV-siMTA2TR#2) or siNC (LV-siNC) were transplanted subcutaneously into nude mice. (A) In vivo fluorescence imaging demonstrated the subcutaneous tumor. (B) The weight of subcutaneous tumor was measured after 4 weeks when mice were sacrificed. (C) The volume of subcutaneous tumor was measured every 4 days. (D) MTA2TR expression was compared in the subcutaneous tumor form LV-siMTA2TR#1, LV-siMTA2TR#2 or LV-siNC group. (E) MTA2 mRNA expression was detected in the subcutaneous tumor form LV-siMTA2TR#1, LV-siMTA2TR#2 or LV-siNC group. (F) The MTA2 protein expression was demonstrated by IHC in the subcutaneous tumor form LV-siMTA2TR#1, LV-siMTA2TR#2 or LV-siNC group. (G-K) BxPC-3 cells transfected with LV-siMTA2TR#1, LV-siMTA2TR#2 or LV-siNC were transplanted via tail vein injection to observe tumor metastasis. (G) In vivo fluorescence imaging indicated the metastasis of BxPC-3 cells in LV-siMTA2TR#1, LV-siMTA2TR#2 or LV-siNC group when mice were sacrificed after 4 weeks. Arrows indicate the invasion nodules. Scale bars, 1 cm. (H) Liver metastasis was measured with the indicated BxPC-3 cells. N=5 mice in each group. (I) The number of visible liver metastases per 5 sections in each nude mouse. (J, K) H&E images of liver and lung tissue isolated from LV-siMTA2TR#1, LV-siMTA2TR#2 or LV-siNC group. Arrows indicate the invasion nodules. Scale bars, 100 μm.

Article Snippet: Protein extracts were denatured, underwent 10% SDS-PAGE gel electrophoresis, and were then transferred to PVDF membranes (Millipore) and probed with the following primary antibodies: MTA2 (Proteintech, 66195-1-Ig), ATF3 (Abcam, ab207434), HIF-1α (Proteintech, 20960-1-AP), PAN-AC (Abconal, A2391), GAPDH (Proteintech, 10494-1-AP), β-actin (Proteintech, 20536-1-AP).

Techniques: In Vivo, Transfection, Fluorescence, Imaging, Expressing, Injection, Isolation

Journal: Theranostics

Article Title: LncRNA-MTA2TR functions as a promoter in pancreatic cancer via driving deacetylation-dependent accumulation of HIF-1α

doi: 10.7150/thno.34559

Figure Lengend Snippet: MTA2TR predominantly functions via regulating MTA2. (A) Schematic annotation the MTA2TR and MTA2 genomic locus. Arrows indicate transcript orientation. (B) Levels of MTA2 mRNA and protein in MTA2TR-knockdown BxPC-3/SW1990 cells were detected via qRT-PCR and Western blot, respectively. (C) BxPC-3 and SW1990 cells were transfected using empty vector as negative control (Vector) or 1.6, 3.2, 6.4, or 12.8μg MTA2TR overexpression vector (MTA2TR-OE). MTA2 mRNA and protein levels were assessed via qRT-PCR and Western blot, respectively. (D, E) The regulation of MTA2TR knockdown or overexpression on MTA2 premRNA expression was detected respectively. (F, G) Total cellular RNA was collected 0, 1, 2, 4, and 6 hours following actinomycin D treatment in MTA2TR knockdown or overexpression BxPC-3 cells and subjected to qRT-PCR. (H) The effect of MTA2TR-knockdown or MTA2TR-overexpression on promoter activity of MTA2 was compared via luciferase reporter assay in BxPC-3 cells co-transfected using plasmids containing wild-type MTA2 promoter.

Article Snippet: Protein extracts were denatured, underwent 10% SDS-PAGE gel electrophoresis, and were then transferred to PVDF membranes (Millipore) and probed with the following primary antibodies: MTA2 (Proteintech, 66195-1-Ig), ATF3 (Abcam, ab207434), HIF-1α (Proteintech, 20960-1-AP), PAN-AC (Abconal, A2391), GAPDH (Proteintech, 10494-1-AP), β-actin (Proteintech, 20536-1-AP).

Techniques: Knockdown, Quantitative RT-PCR, Western Blot, Transfection, Plasmid Preparation, Negative Control, Over Expression, Expressing, Activity Assay, Luciferase, Reporter Assay

Journal: Theranostics

Article Title: LncRNA-MTA2TR functions as a promoter in pancreatic cancer via driving deacetylation-dependent accumulation of HIF-1α

doi: 10.7150/thno.34559

Figure Lengend Snippet: MTA2TR is associated with transcription factor ATF3. (A) Venn diagram of the overlap indicated ATF3 as a MTA2TR/MTA2-associated protein from ChIPBase, JASPAR, and catRAPID. (B) Schematic showing ATF3 binding sites (P1, P2) on the MTA2 promoter. (C) The correlation between ATF3 and MTA2 expression was assessed in 183 PC samples from ChIPBase. (D) Schematic illustration of association between MTA2TR and ATF3. Red coloration indicates stronger confidence for a given base. (E) The binding between MTA2TR and ATF3 was verified by RIP assay with anti-ATF3 antibody in BxPC-3/SW1990 cells, and the co-precipitated transcript were determined using qRT-PCR. Positive Control Antibody, the Anti-SNRNP70 rabbit polyclonal antibody, is expected by sequence similarity of the immunogen to cross react with SNRNP70 of human, mouse, rat and canine origins. The U1 snRNA RIP primers should also be able to amplify U1 snRNA converted cDNA in those species. (F) The binding between MTA2TR and ATF3 was verified by RNA-pulldown assay with a biotinylated MTA2TR probe in BxPC-3/SW1990 cells. The immunoblot analysis showed the pulldown protein with anti-ATF3 and anti-HuR antibody. RNA from the 3´ untranslated-region (UTR) of the androgen receptor (AR) served as the control, containing UC-rich HuR binding regions. MTA2TR-antisense, control RNA, lacked ATF3 or HuR binding sites. (G) MTA2TR deletion mutants were transcribed in vitro and applied for RNA pull-down assays. (H) After transfected with empty vector and plasmids containing p1, p2, p3 fragment, MTA2 mRNA and protein levels were assessed. (I) BxPC-3/SW1990 cells were transfected using empty vector as negative control (Vector) or 1.6, 3.2, 6.4, or 12.8μg P2-containing overexpression vector. MTA2 protein levels were measured.

Article Snippet: Protein extracts were denatured, underwent 10% SDS-PAGE gel electrophoresis, and were then transferred to PVDF membranes (Millipore) and probed with the following primary antibodies: MTA2 (Proteintech, 66195-1-Ig), ATF3 (Abcam, ab207434), HIF-1α (Proteintech, 20960-1-AP), PAN-AC (Abconal, A2391), GAPDH (Proteintech, 10494-1-AP), β-actin (Proteintech, 20536-1-AP).

Techniques: Binding Assay, Expressing, Quantitative RT-PCR, Positive Control, Sequencing, Western Blot, Control, In Vitro, Transfection, Plasmid Preparation, Negative Control, Over Expression

Journal: Theranostics

Article Title: LncRNA-MTA2TR functions as a promoter in pancreatic cancer via driving deacetylation-dependent accumulation of HIF-1α

doi: 10.7150/thno.34559

Figure Lengend Snippet: MTA2TR regulates MTA2 transcription via recruiting ATF3 to the MTA2 promoter. (A, B) MTA2 mRNA and protein levels were detected in ATF3-knockdown or ATF3-overexpression pancreatic cancer cells, respectively. (C) The binding of ATF3 on MTA2 promoter region (P1, P2) was verified with ChIP assay putative binding sites of ATF3 on the MTA2 promoter in BxPC-3 cells. (D) The vector containing WT and three mutants of ATF3 binding sites on the MTA2 promoter were designed for the luciferase reporter assay. (E) The effect of ATF3 on the MTA2 promoter activity was analyzed following transfection of BxPC-3 cells using a WT- or MUT-containing reporter vector. (F) MTA2 mRNA and protein levels were detected in MTA2TR-overexpression or/and ATF3-knockdown PC cells. (G) The binding between ATF3 and MTA2 promoter was evaluated by ChIP assay in MTA2TR-overexpression or/and ATF3-knockdown BxPC-3 cells. (H) The activity of MTA2 promoter was determined with a luciferase reporter assay in MTA2TR-overexpression or/and ATF3-knockdown BxPC-3 cells.

Article Snippet: Protein extracts were denatured, underwent 10% SDS-PAGE gel electrophoresis, and were then transferred to PVDF membranes (Millipore) and probed with the following primary antibodies: MTA2 (Proteintech, 66195-1-Ig), ATF3 (Abcam, ab207434), HIF-1α (Proteintech, 20960-1-AP), PAN-AC (Abconal, A2391), GAPDH (Proteintech, 10494-1-AP), β-actin (Proteintech, 20536-1-AP).

Techniques: Knockdown, Over Expression, Binding Assay, Plasmid Preparation, Luciferase, Reporter Assay, Activity Assay, Transfection

Journal: Theranostics

Article Title: LncRNA-MTA2TR functions as a promoter in pancreatic cancer via driving deacetylation-dependent accumulation of HIF-1α

doi: 10.7150/thno.34559

Figure Lengend Snippet: MTA2TR is upregulated during hypoxia condition. (A) MTA2 expression was assessed in BxPC-3/SW1990 cells during hypoxia with different time point. (B) MTA2TR expression was assessed in HIF-1α-knockdown BxPC-3/SW1990 cells under hypoxia condition. (C) The MTA2TR expression in HIF-1α-knockdown BxPC-3 cells was shown by RNA-FISH under normoxia or hypoxia condition. Nucleus was counterstained with DAPI. Scale bars: 20μm. (D) The putative HREs (P1, P2, and P3) on the MTA2TR promoter area was shown. (E) HIF-1α binding to the MTA2TR promoter was verified by ChIP assay in BxPC-3 cells. (F) How hypoxia affects HIF-1α binding to the MTA2TR promoter was inspected by ChIP assay in BxPC-3 cells. (G) Following transfection of BxPC-3 cells using a vector containing WT- and MUT-HRE of MTA2TR promoter, MTA2TR promoter activity was analyzed in the reporter cells co-transfected with siHIF-1α#1 under normoxia or hypoxia condition.

Article Snippet: Protein extracts were denatured, underwent 10% SDS-PAGE gel electrophoresis, and were then transferred to PVDF membranes (Millipore) and probed with the following primary antibodies: MTA2 (Proteintech, 66195-1-Ig), ATF3 (Abcam, ab207434), HIF-1α (Proteintech, 20960-1-AP), PAN-AC (Abconal, A2391), GAPDH (Proteintech, 10494-1-AP), β-actin (Proteintech, 20536-1-AP).

Techniques: Expressing, Knockdown, Binding Assay, Transfection, Plasmid Preparation, Activity Assay

Journal: Theranostics

Article Title: LncRNA-MTA2TR functions as a promoter in pancreatic cancer via driving deacetylation-dependent accumulation of HIF-1α

doi: 10.7150/thno.34559

Figure Lengend Snippet: Hypoxia-induced MTA2TR stabilizes HIF-1α by promoting deacetylation. (A) HIF-1α mRNA and protein levels were analyzed in MTA2TR-knockdown BxPC-3/SW1990 cells. (B) The effect of MTA2TR knockdown on stabilization of HIF-1α was measured in BxPC-3/SW1990 cells treated with cycloheximide under hypoxia for the indicated periods of time. (C) HIF-1α levels in the MTA2TR-knockdown BxPC-3/SW1990 cells was detected under hypoxia with or without MG132 treatment. (D, E) HIF-1α acetylation and the binding between MTA2 and HIF-1α in the MTA2TR-knockdown BxPC-3/SW1990 cells treated with 1% O 2 24 h were analyzed in the cell lysates following anti-HIF-1α or anti-MTA2 immunoprecipitation. (F) The acetylation of HIF-1α and the binding between MTA2 and HIF-1α in the BxPC-3/SW1990 cells, cultured under hypoxia condition, transfected with siMTA2TR#1 or/and MTA2-OE were analyzed by IP with anti-HIF-1α or anti-MTA2 antibody. (G) The mRNA level of VEGFa, VEGFb, LOX, LOXL2, PLOD2 and TUBA1 was detected in MTA2TR-knockdown BxPC-3 and SW1990 cells under hypoxia condition by qRT-PCR.

Article Snippet: Protein extracts were denatured, underwent 10% SDS-PAGE gel electrophoresis, and were then transferred to PVDF membranes (Millipore) and probed with the following primary antibodies: MTA2 (Proteintech, 66195-1-Ig), ATF3 (Abcam, ab207434), HIF-1α (Proteintech, 20960-1-AP), PAN-AC (Abconal, A2391), GAPDH (Proteintech, 10494-1-AP), β-actin (Proteintech, 20536-1-AP).

Techniques: Knockdown, Binding Assay, Immunoprecipitation, Cell Culture, Transfection, Quantitative RT-PCR

Journal: Theranostics

Article Title: LncRNA-MTA2TR functions as a promoter in pancreatic cancer via driving deacetylation-dependent accumulation of HIF-1α

doi: 10.7150/thno.34559

Figure Lengend Snippet: MTA2TR is associated with the hypoxia-induced HIF-1α/MTA2 pathway. (A) The representative immunofluorescence of MTA2 and HIF-1α in BxPC-3 cells after transfection using siMTA2TR#1, siMTA2TR#2, siMTA2#1, or siMTA2#2 under normoxia or hypoxic conditions. (B) The representative immunofluorescence of MTA2 and HIF-1α in BxPC-3 cells following co-transfection using siHIF-1α#1, siHIF-1α#2 and MTA2TR-OE or MTA2-OE under hypoxia conditions. Scale bars: 50 μm.

Article Snippet: Protein extracts were denatured, underwent 10% SDS-PAGE gel electrophoresis, and were then transferred to PVDF membranes (Millipore) and probed with the following primary antibodies: MTA2 (Proteintech, 66195-1-Ig), ATF3 (Abcam, ab207434), HIF-1α (Proteintech, 20960-1-AP), PAN-AC (Abconal, A2391), GAPDH (Proteintech, 10494-1-AP), β-actin (Proteintech, 20536-1-AP).

Techniques: Immunofluorescence, Transfection, Cotransfection

Journal: Theranostics

Article Title: LncRNA-MTA2TR functions as a promoter in pancreatic cancer via driving deacetylation-dependent accumulation of HIF-1α

doi: 10.7150/thno.34559

Figure Lengend Snippet: Correlation of MTA2TR, ATF3, MTA2 and HIF-1α expression in PC samples. (A) Kaplan-Meier curves showing overall survivals in 40 cases of PC. PC patients with MTA2TR expression level below or above the 50th percentile was classified as low or high MTA2TR group. (B) The Pearson's correlation between MTA2TR and MTA2 expression was analyzed in 40 PC tissues. (C) Kaplan-Meier curves for overall survival of 114 PC patients from OncoLnc database. MTA2 expression level below or above the 50th percentile was classified as low or high MTA2 group. (D) Combined immunofluorescence of HIF-1α protein (green) and RNA-FISH analysis of MTA2TR (red) in PC and corresponding NP tissues. Scale bars, 50 μm. (E) The correlation between HIF-1α and MTA2 expression was assessed in 183 PC samples from ChIPBase. (F) Graphical representation of the regulation and function of MTA2TR in PC.

Article Snippet: Protein extracts were denatured, underwent 10% SDS-PAGE gel electrophoresis, and were then transferred to PVDF membranes (Millipore) and probed with the following primary antibodies: MTA2 (Proteintech, 66195-1-Ig), ATF3 (Abcam, ab207434), HIF-1α (Proteintech, 20960-1-AP), PAN-AC (Abconal, A2391), GAPDH (Proteintech, 10494-1-AP), β-actin (Proteintech, 20536-1-AP).

Techniques: Expressing, Immunofluorescence

Journal: Oncotarget

Article Title: MTA1 promotes epithelial to mesenchymal transition and metastasis in non-small-cell lung cancer

doi: 10.18632/oncotarget.16404

Figure Lengend Snippet: (A) Western blot(left) and RT-PCR (right) analyses of MTA1 expression in four cell lines. ** P <0.01, *** P <0.001, compared with Beas-2b cells. GAPDH was used as loading control. Student t-test was used for statistical analyze for three independent experiments. For PCR amplification, 25 cycles were performed. (B) Immunofluorescence staining for MTA1 in four cell lines (red: MTA1; blue: nuclei). (C) Western blot analysis of MTA1 expression in MTA1-overexpressing cells, MTA1-silenced cells, and control cells. GAPDH was used as a loading control. oex: overexpression; sh#1: shRNA#1.

Article Snippet: The following primary antibodies were used: MTA1 (Cell Signaling, #5647, 1:1,000), E-cadherin (Proteintech, 20874-1-AP, 1:1,000), Vimentin (Proteintech, 10366-1-AP, 1:2,000), Claudin-1 (Cell Signaling, #13255, 1:1,000), ZO-1 (Cell Signaling, #8193, 1:1,000), p-AKT (WLP001a, 1:750), AKT (WL0003b, 1:750), p-GSK3β (Cell Signaling, #9323,1:1,000), GSK3β (Cell Signaling #9315, 1:1,000), β-catenin (Proteintech, 51067-2-AP, 1:1,000), GAPDH (HRP-60004, 1:5,000), and Wnt1 (Sangon Biotech, D261302, 1:50) Secondary antibody (Cell Signaling, #7044) was diluted 1:2,000.

Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction, Expressing, Control, Amplification, Immunofluorescence, Staining, Over Expression, shRNA

Journal: Oncotarget

Article Title: MTA1 promotes epithelial to mesenchymal transition and metastasis in non-small-cell lung cancer

doi: 10.18632/oncotarget.16404

Figure Lengend Snippet: (A) Western blot analysis of MTA1, E-cadherin, Vimentin, Claudin-1, and ZO-1 expression GAPDH was used as a loading control. Quantitative data are shown in (B–F) . Student t-test was used for statistical analyze for three independent experiments. * P <0.05, ** P <0.01, *** P <0.001. (G) Cell cytoskeleton images were obtained via confocal microscopy MTA1 overexpression changed cells from relatively circular to irregular in shape, with the emergence of prolonged “feet.” The “feet” of MTA1-shRNA-treated cells were shortened, and cells changed from long spindles to relatively elliptical or circular in shape compared with controls. Red: F-actin; blue: nuclei (bars: 50 μm). →: representative changes; oex: overexpression; sh#1: shRNA#1.

Article Snippet: The following primary antibodies were used: MTA1 (Cell Signaling, #5647, 1:1,000), E-cadherin (Proteintech, 20874-1-AP, 1:1,000), Vimentin (Proteintech, 10366-1-AP, 1:2,000), Claudin-1 (Cell Signaling, #13255, 1:1,000), ZO-1 (Cell Signaling, #8193, 1:1,000), p-AKT (WLP001a, 1:750), AKT (WL0003b, 1:750), p-GSK3β (Cell Signaling, #9323,1:1,000), GSK3β (Cell Signaling #9315, 1:1,000), β-catenin (Proteintech, 51067-2-AP, 1:1,000), GAPDH (HRP-60004, 1:5,000), and Wnt1 (Sangon Biotech, D261302, 1:50) Secondary antibody (Cell Signaling, #7044) was diluted 1:2,000.

Techniques: Western Blot, Expressing, Control, Confocal Microscopy, Over Expression, shRNA

Journal: Oncotarget

Article Title: MTA1 promotes epithelial to mesenchymal transition and metastasis in non-small-cell lung cancer

doi: 10.18632/oncotarget.16404

Figure Lengend Snippet: Cells were transfected with MTA1-overexpression plasmid or infected with lentivirus containing shRNA for 72 h, and then were collected to perform the following experiments. Quantified data (A) and representative images (B) of migration (left) and invasion assays (right) in Beas-2b and H460 cells. Quantified data (C) and representative images (D) of migration (left) and invasion assays (right) in Beas-2b and H460 cells. Student t-test was used for statistical analyze for three independent experiments. * P <0.05, ** P <0.01. (E) Western blot analysis of MTA1, E-cadherin, Vimentin, Claudin-1, and ZO-1 expression GAPDH was used as a loading control. oex: overexpression, sh#1: shRNA#1, sh#2: shRNA#2, sh#3: shRNA#3.

Article Snippet: The following primary antibodies were used: MTA1 (Cell Signaling, #5647, 1:1,000), E-cadherin (Proteintech, 20874-1-AP, 1:1,000), Vimentin (Proteintech, 10366-1-AP, 1:2,000), Claudin-1 (Cell Signaling, #13255, 1:1,000), ZO-1 (Cell Signaling, #8193, 1:1,000), p-AKT (WLP001a, 1:750), AKT (WL0003b, 1:750), p-GSK3β (Cell Signaling, #9323,1:1,000), GSK3β (Cell Signaling #9315, 1:1,000), β-catenin (Proteintech, 51067-2-AP, 1:1,000), GAPDH (HRP-60004, 1:5,000), and Wnt1 (Sangon Biotech, D261302, 1:50) Secondary antibody (Cell Signaling, #7044) was diluted 1:2,000.

Techniques: Transfection, Over Expression, Plasmid Preparation, Infection, shRNA, Migration, Western Blot, Expressing, Control

Journal: Oncotarget

Article Title: MTA1 promotes epithelial to mesenchymal transition and metastasis in non-small-cell lung cancer

doi: 10.18632/oncotarget.16404

Figure Lengend Snippet: (A) Lung metastasis in mice after intravenous tail vail injection of cancer cells Bars: 5.0 mm. →: Suspicious metastasis lesions. (B) HE staining of lung sections Bars: 250μm. →: Confirmed metastasis lesions. (C) Quantitative analysis(n=5) of lung metastasis tumor volume, calculated when bars=250μm. (D) Immunohistochemical staining for MTA1, E-cadherin, Vimentin, Claudin-1, and ZO-1 in lung tissues. Bars=50μm. (E) Expression intensity (MTA1, E-cadherin, Vimentin, Claudin-1, ZO-1) was calculated as IOD/Area (n=5). oex: overexpression; sh#1: shRNA#1. Student t-test was used for statistical analyze for three independent experiments. NS: no significance, P >0.05. * P <0.05, ** P <0.01, *** P <0.001.

Article Snippet: The following primary antibodies were used: MTA1 (Cell Signaling, #5647, 1:1,000), E-cadherin (Proteintech, 20874-1-AP, 1:1,000), Vimentin (Proteintech, 10366-1-AP, 1:2,000), Claudin-1 (Cell Signaling, #13255, 1:1,000), ZO-1 (Cell Signaling, #8193, 1:1,000), p-AKT (WLP001a, 1:750), AKT (WL0003b, 1:750), p-GSK3β (Cell Signaling, #9323,1:1,000), GSK3β (Cell Signaling #9315, 1:1,000), β-catenin (Proteintech, 51067-2-AP, 1:1,000), GAPDH (HRP-60004, 1:5,000), and Wnt1 (Sangon Biotech, D261302, 1:50) Secondary antibody (Cell Signaling, #7044) was diluted 1:2,000.

Techniques: Injection, Staining, Immunohistochemical staining, Expressing, Over Expression, shRNA

Journal: Oncotarget

Article Title: MTA1 promotes epithelial to mesenchymal transition and metastasis in non-small-cell lung cancer

doi: 10.18632/oncotarget.16404

Figure Lengend Snippet: (A) Western blot analysis of AKT, p-AKT, GSK-3β, p-GSK-3β, and β-catenin expression, (B) Western blot analysis of Wnt1. (C) Western blot analysis of MTA1, AKT, p-AKT, GSK-3β, p-GSK-3β, β-catenin, and EMT markers (E-cadherin, Vimentin, Claudin-1, and ZO-1). Cells were treated with 10 μM MK2206 for 24 h. (D) Western blot analysis of MTA1, AKT, p-AKT, GSK-3β, p-GSK-3β, β-catenin, and EMT markers (E-cadherin, Vimentin, Claudin-1, and ZO-1). H460 cells were transfected with MTA1-overexpression plasmid for 72 h, or transfected with siRNA for 48 h, or transfected with MTA1-overexpression plasmid for 24 h followed by siRNA for 48 h. GAPDH was used as a loading control. oex: overexpression; si#1: siRNA#1; si#2: siRNA#2.

Article Snippet: The following primary antibodies were used: MTA1 (Cell Signaling, #5647, 1:1,000), E-cadherin (Proteintech, 20874-1-AP, 1:1,000), Vimentin (Proteintech, 10366-1-AP, 1:2,000), Claudin-1 (Cell Signaling, #13255, 1:1,000), ZO-1 (Cell Signaling, #8193, 1:1,000), p-AKT (WLP001a, 1:750), AKT (WL0003b, 1:750), p-GSK3β (Cell Signaling, #9323,1:1,000), GSK3β (Cell Signaling #9315, 1:1,000), β-catenin (Proteintech, 51067-2-AP, 1:1,000), GAPDH (HRP-60004, 1:5,000), and Wnt1 (Sangon Biotech, D261302, 1:50) Secondary antibody (Cell Signaling, #7044) was diluted 1:2,000.

Techniques: Western Blot, Expressing, Transfection, Over Expression, Plasmid Preparation, Control

Journal: Oncotarget

Article Title: MTA1 promotes epithelial to mesenchymal transition and metastasis in non-small-cell lung cancer

doi: 10.18632/oncotarget.16404

Figure Lengend Snippet: (A) Representative immunohistochemical staining for MTA1 in NSCLC and normal lung tissues Top to bottom: normal lung; negative immunostaining in NSCLC tissues; light in NSCLC tissues; moderate in NSCLC tissues; intense in NSCLC tissues. Bars: 50μm. (B) Kaplan-Meier survival curves for MTA1 expression in 54 NSCLC patients Relationships between MTA1 expression and p-AKT and β-catenin expression in NSCLC tissues. (C) Representative immunohistochemical staining for MTA1, p-AKT, and β-catenin. Bars: 50μm.

Article Snippet: The following primary antibodies were used: MTA1 (Cell Signaling, #5647, 1:1,000), E-cadherin (Proteintech, 20874-1-AP, 1:1,000), Vimentin (Proteintech, 10366-1-AP, 1:2,000), Claudin-1 (Cell Signaling, #13255, 1:1,000), ZO-1 (Cell Signaling, #8193, 1:1,000), p-AKT (WLP001a, 1:750), AKT (WL0003b, 1:750), p-GSK3β (Cell Signaling, #9323,1:1,000), GSK3β (Cell Signaling #9315, 1:1,000), β-catenin (Proteintech, 51067-2-AP, 1:1,000), GAPDH (HRP-60004, 1:5,000), and Wnt1 (Sangon Biotech, D261302, 1:50) Secondary antibody (Cell Signaling, #7044) was diluted 1:2,000.

Techniques: Immunohistochemical staining, Staining, Immunostaining, Expressing

Journal: Oncotarget

Article Title: MTA1 promotes epithelial to mesenchymal transition and metastasis in non-small-cell lung cancer

doi: 10.18632/oncotarget.16404

Figure Lengend Snippet: Associations between MTA1 expression and NSCLC patient clinicopathological characteristics

Article Snippet: The following primary antibodies were used: MTA1 (Cell Signaling, #5647, 1:1,000), E-cadherin (Proteintech, 20874-1-AP, 1:1,000), Vimentin (Proteintech, 10366-1-AP, 1:2,000), Claudin-1 (Cell Signaling, #13255, 1:1,000), ZO-1 (Cell Signaling, #8193, 1:1,000), p-AKT (WLP001a, 1:750), AKT (WL0003b, 1:750), p-GSK3β (Cell Signaling, #9323,1:1,000), GSK3β (Cell Signaling #9315, 1:1,000), β-catenin (Proteintech, 51067-2-AP, 1:1,000), GAPDH (HRP-60004, 1:5,000), and Wnt1 (Sangon Biotech, D261302, 1:50) Secondary antibody (Cell Signaling, #7044) was diluted 1:2,000.

Techniques: Expressing

Journal: Oncotarget

Article Title: MTA1 promotes epithelial to mesenchymal transition and metastasis in non-small-cell lung cancer

doi: 10.18632/oncotarget.16404

Figure Lengend Snippet: Univariate analysis of NSCLC patient prognosis (n=54)

Article Snippet: The following primary antibodies were used: MTA1 (Cell Signaling, #5647, 1:1,000), E-cadherin (Proteintech, 20874-1-AP, 1:1,000), Vimentin (Proteintech, 10366-1-AP, 1:2,000), Claudin-1 (Cell Signaling, #13255, 1:1,000), ZO-1 (Cell Signaling, #8193, 1:1,000), p-AKT (WLP001a, 1:750), AKT (WL0003b, 1:750), p-GSK3β (Cell Signaling, #9323,1:1,000), GSK3β (Cell Signaling #9315, 1:1,000), β-catenin (Proteintech, 51067-2-AP, 1:1,000), GAPDH (HRP-60004, 1:5,000), and Wnt1 (Sangon Biotech, D261302, 1:50) Secondary antibody (Cell Signaling, #7044) was diluted 1:2,000.

Techniques: Biomarker Discovery, Expressing

Journal: Oncotarget

Article Title: MTA1 promotes epithelial to mesenchymal transition and metastasis in non-small-cell lung cancer

doi: 10.18632/oncotarget.16404

Figure Lengend Snippet: Multivariate analysis of NSCLC patient prognosis (n=54)

Article Snippet: The following primary antibodies were used: MTA1 (Cell Signaling, #5647, 1:1,000), E-cadherin (Proteintech, 20874-1-AP, 1:1,000), Vimentin (Proteintech, 10366-1-AP, 1:2,000), Claudin-1 (Cell Signaling, #13255, 1:1,000), ZO-1 (Cell Signaling, #8193, 1:1,000), p-AKT (WLP001a, 1:750), AKT (WL0003b, 1:750), p-GSK3β (Cell Signaling, #9323,1:1,000), GSK3β (Cell Signaling #9315, 1:1,000), β-catenin (Proteintech, 51067-2-AP, 1:1,000), GAPDH (HRP-60004, 1:5,000), and Wnt1 (Sangon Biotech, D261302, 1:50) Secondary antibody (Cell Signaling, #7044) was diluted 1:2,000.

Techniques: Expressing

Journal: Oncotarget

Article Title: MTA1 promotes epithelial to mesenchymal transition and metastasis in non-small-cell lung cancer

doi: 10.18632/oncotarget.16404

Figure Lengend Snippet: Correlations between MTA1 expression and p-AKT or β-catenin expression in NSCLC tissues

Article Snippet: The following primary antibodies were used: MTA1 (Cell Signaling, #5647, 1:1,000), E-cadherin (Proteintech, 20874-1-AP, 1:1,000), Vimentin (Proteintech, 10366-1-AP, 1:2,000), Claudin-1 (Cell Signaling, #13255, 1:1,000), ZO-1 (Cell Signaling, #8193, 1:1,000), p-AKT (WLP001a, 1:750), AKT (WL0003b, 1:750), p-GSK3β (Cell Signaling, #9323,1:1,000), GSK3β (Cell Signaling #9315, 1:1,000), β-catenin (Proteintech, 51067-2-AP, 1:1,000), GAPDH (HRP-60004, 1:5,000), and Wnt1 (Sangon Biotech, D261302, 1:50) Secondary antibody (Cell Signaling, #7044) was diluted 1:2,000.

Techniques: Expressing