primary anti twist1  (Boster Bio)

Journal: Burns & Trauma

Article Title: Twist-related protein 1 promotes transforming growth factor β receptor 1 in keloid fibroblasts via regulating the stability of myocyte enhancer factor 2A

doi: 10.1093/burnst/tkae024

Figure Lengend Snippet: TWIST1 is upregulated in keloid tissue and keloid-derived fibroblasts. ( a ) The expression levels of TWIST1 were detected in keloid and paired normal tissue by immunohistochemistry (Scale bar: 100 µm & 50 µm). ( b ) mRNA expression level of TWIST1 was measured in keloid and normal skin tissue by real-time PCR. ( c , d ) Western blotting was used to measure the protein expression level of TWIST1 keloid tissues and paired normal skin tissues with densitometry analysis of western blot results. ( e , f ) Western blot results of TWIST1 in keloid-derived fibroblasts and normal skin-derived fibroblasts. ( g ) Western blot results of TWIST1 in keloid-derived fibroblasts with 0, 5 and 10 ng/ml TGF-β1. * p < 0.05, * * p < 0.01, * * * p < 0.001. NFBs normal skin-derived fibroblasts, KFBs keloid-derived fibroblasts, TWIST1 Twist-related protein 1, TGF-β transforming growth factor beta

Article Snippet: Sections were stained using an anti-TWIST1 primary antibody (CST, 69366, 1 : 300) and anti-MEF2A primary antibody (Santa Cruz, sc-17 785, 1 : 300) for immunohistochemistry at 4°C overnight.

Techniques: Derivative Assay, Expressing, Immunohistochemistry, Real-time Polymerase Chain Reaction, Western Blot

Journal: Burns & Trauma

Article Title: Twist-related protein 1 promotes transforming growth factor β receptor 1 in keloid fibroblasts via regulating the stability of myocyte enhancer factor 2A

doi: 10.1093/burnst/tkae024

Figure Lengend Snippet: Knockdown and overexpression of TWIST1 affect the proliferation and invasion of keloid-derived fibroblasts. ( a ) Proliferation of keloid-derived fibroblasts in small interfering negative control and small interfering TWIST1 groups was detected by the cell counting kit-8 assay. ( b ) Proliferation of keloid-derived fibroblasts in vector and TWIST1 groups was detected by cell counting kit-8 assay. ( c ) Invasion ability of keloid-derived fibroblasts in small interfering negative control and small interfering TWIST1 groups was detected by transwell assays (Scale bar: 100 µm). ( d ) Invasion of keloid-derived fibroblasts in vector and TWIST1 groups was detected by transwell assays (Scale bar: 100 µm). ( e ) Immunofluorescence assay of α-SMA in keloid-derived fibroblasts with 5 ng/ml TGF-β1 (Scale bar: 50 µm). * * p < 0.01, * * * p < 0.001. TWIST1 Twist-related protein 1, TGFβ1 transforming growth factor 1, si small interfering, OD optical density, OE over-expression, NC negative control

Article Snippet: Sections were stained using an anti-TWIST1 primary antibody (CST, 69366, 1 : 300) and anti-MEF2A primary antibody (Santa Cruz, sc-17 785, 1 : 300) for immunohistochemistry at 4°C overnight.

Techniques: Knockdown, Over Expression, Derivative Assay, Negative Control, Cell Counting, Plasmid Preparation, Immunofluorescence

Journal: Burns & Trauma

Article Title: Twist-related protein 1 promotes transforming growth factor β receptor 1 in keloid fibroblasts via regulating the stability of myocyte enhancer factor 2A

doi: 10.1093/burnst/tkae024

Figure Lengend Snippet: TWIST1 knockdown and overexpression affect the expression of the TGFβ/Smad signaling-related molecules TΒR1, Smad2 and Smad3, and the fibrosis markers α-SMA, COL1 and COL3. ( a ) Protein expression of TβRI, TβRII and Smad3, and fibrosis markers α-SMA, COL1 and COL3 in small interfering negative control and small interfering TWIST1 groups was detected by western blotting. ( b ) Protein expression of TβRI, TβRII and Smad3 and the fibrosis markers α-SMA, COL1 and COL3 in vector and TWIST1 groups was detected by western blotting. ( c , d ) Densitometry analysis of western blot results. * * p < 0.01, * * * p < 0.001; ns not significant. TβR1 transforming growth factor beta receptor 1, SMAD3 SMAD family member 3, COL1 collagen type I, COL3 collagen type III, NC negative control

Article Snippet: Sections were stained using an anti-TWIST1 primary antibody (CST, 69366, 1 : 300) and anti-MEF2A primary antibody (Santa Cruz, sc-17 785, 1 : 300) for immunohistochemistry at 4°C overnight.

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

Journal: Burns & Trauma

Article Title: Twist-related protein 1 promotes transforming growth factor β receptor 1 in keloid fibroblasts via regulating the stability of myocyte enhancer factor 2A

doi: 10.1093/burnst/tkae024

Figure Lengend Snippet: TWIST1 interacts with MEF2A and regulates MEF2A expression. ( a , b ) Cell lysates of keloid-derived fibroblasts were immunoprecipitated with anti-TWIST1 antibody, followed by immunoblotting with anti-TβRI or anti-MEF2A antibody, respectively. ( c ) Cell lysates of keloid-derived fibroblasts were immunoprecipitated with anti-MEF2A antibody, followed by immunoblotting with anti-TWIST1 and anti-MEF2A antibody; IgG was used as a negative control. ( d ) Protein expression of MEF2A in small interfering negative control and small interfering TWIST1 groups was detected by western blotting with densitometry analysis of western blot results. ( e ). Protein expression of MEF2A in vector and TWIST1 groups was detected by western blotting with densitometry analysis of western blot results. * * p < 0.01, * * * p < 0.001. IgG immunoglobulin G, TWIST1 Twist-related protein 1, MEF2A myocyte enhancer factor 2A, NC negative control

Article Snippet: Sections were stained using an anti-TWIST1 primary antibody (CST, 69366, 1 : 300) and anti-MEF2A primary antibody (Santa Cruz, sc-17 785, 1 : 300) for immunohistochemistry at 4°C overnight.

Techniques: Expressing, Derivative Assay, Immunoprecipitation, Western Blot, Negative Control, Plasmid Preparation

Journal: Burns & Trauma

Article Title: Twist-related protein 1 promotes transforming growth factor β receptor 1 in keloid fibroblasts via regulating the stability of myocyte enhancer factor 2A

doi: 10.1093/burnst/tkae024

Figure Lengend Snippet: TWIST1 suppress ubiquitination and degradation of MEF2A. ( a ) Western blotting detected the alteration of MEF2A in small interfering negative control and small interfering TWIST1 groups treated with 10 μg/ml cycloheximide (CHX) for the indicated times. ( b ) Densitometric analysis of MEF2A from experiments. ( c ) Keloid-derived fibroblasts transfected by si-NC or si-TWIST1 were treated with 10 nmol/ml MG132, and protein concentrations of MEF2A were determined. ( d ) Densitometric analysis of MEF2A from experiments. ( e , f ) Lysates from keloid-derived fibroblasts modulated with down- or up-regulated TWIST1 were incubated with anti-MEF2A antibody. Immunoprecipitants were immunoblotted with an anti-Ub (ubiquitin) antibody. ( g ) Coimmunoprecipitation of MDM2 and MEF2A in keloid-derived fibroblasts transfected with si-NC or si-TWIST1. ( h , i ) Protein concentrations of MDM2 in keloid-derived fibroblasts transfected with si-NC or si-TWIST1. * P < 0.05, * * * p < 0.001; ns not significant. Ub ubiquitin, TWIST1 Twist-related protein 1, MEF2A myocyte enhancer factor 2A, NC negative control, CHX cycloheximide, MDM2 mouse double minute 2 homolog, si small interfering

Article Snippet: Sections were stained using an anti-TWIST1 primary antibody (CST, 69366, 1 : 300) and anti-MEF2A primary antibody (Santa Cruz, sc-17 785, 1 : 300) for immunohistochemistry at 4°C overnight.

Techniques: Ubiquitin Proteomics, Western Blot, Negative Control, Derivative Assay, Transfection, Incubation

Journal: Burns & Trauma

Article Title: Twist-related protein 1 promotes transforming growth factor β receptor 1 in keloid fibroblasts via regulating the stability of myocyte enhancer factor 2A

doi: 10.1093/burnst/tkae024

Figure Lengend Snippet: TWIST1 promoted a profibrotic phenotype via the MEF2A-dependent TGF-β pathway in keloid derived fibroblasts. ( a ) Proliferation ability of keloid-derived fibroblasts transfected by control or TWIST1 or TWIST1 with si-MEF2A was measured using a growth curve. ( b ) Cell invasion determined by transwell in pretreated keloid-derived fibroblasts (Scale bar: 100 µm). ( c ) Keloid-derived fibroblasts transfected by control or TWIST1 or TWIST1 with si-MEF2A pretreated with 5 ng/ml TGF-β1 were measured by α-SMA using an immunofluorescence assay (Scale bar: 100 µm). ( d , e ) Protein levels of TβRI, Smad3 and α-SMA were detected by Western blotting. Vinculin was used as internal control. * p < 0.05, * * p < 0.01, * * * p < 0.001. TWIST1 Twist-related protein 1, MEF2A myocyte enhancer factor 2A, NC negative control, TβR1 transforming growth factor beta receptor 1, SMAD3 SMAD family member 3, COL1 collagen type I, COL3 collagen type III

Article Snippet: Sections were stained using an anti-TWIST1 primary antibody (CST, 69366, 1 : 300) and anti-MEF2A primary antibody (Santa Cruz, sc-17 785, 1 : 300) for immunohistochemistry at 4°C overnight.

Techniques: Derivative Assay, Transfection, Control, Immunofluorescence, Western Blot, Negative Control

Journal: Burns & Trauma

Article Title: Twist-related protein 1 promotes transforming growth factor β receptor 1 in keloid fibroblasts via regulating the stability of myocyte enhancer factor 2A

doi: 10.1093/burnst/tkae024

Figure Lengend Snippet: Schematic representation of TWIST1 promoting expression of TGF-β receptor 1 by regulating the stability of MEF2A. TWIST1 Twist-related protein 1, MEF2A myocyte enhancer factor 2A, TβR1 transforming growth factor beta receptor 1, Ub ubiquitin

Article Snippet: Sections were stained using an anti-TWIST1 primary antibody (CST, 69366, 1 : 300) and anti-MEF2A primary antibody (Santa Cruz, sc-17 785, 1 : 300) for immunohistochemistry at 4°C overnight.

Techniques: Expressing, Ubiquitin Proteomics

Journal: Bioengineered

Article Title: CircRAB3IP upregulates twist family BHLH transcription factor (TWIST1) to promote osteosarcoma progression by sponging miR-580-3p.

doi: 10.1080/21655979.2021.1948487

Figure Lengend Snippet: Figure 4. TWIST1 was targeted by miR-580-3p in OS (a) Bioinformatics was used to predict the potential binding site between miR- 580-3p and TWIST1. (b) TWIST1 was a target gene of miR-580-3p. (c, d) miR-580-3p mimics inhibited TWIST1 expression. (e) qRT-PCR indicated that TWIST1 was up-regulated in OS tissues. (f) Pearson’s analysis showed that circRAB3IP expression was positively correlated with TWIST1 expression in OS tissues. OS cells were transfected with sh-NC, sh-circRAB3IP, sh-circRAB3IP+miR-580-3p inhibitor. (g) Luciferase reporter assay was used to determined the activity of TWIST1-Wt reporter. (h) TWIST1 expression was detected by Western blot. *P < 0.05.

Article Snippet: Primary rabbit antihuman antibodies against TWIST1 (1:1000, cell signaling technology, #46702) and GAPDH (1:2000, Abcam, ab9485) were incubated overnight at 4°C.

Techniques: Binding Assay, Expressing, Quantitative RT-PCR, Transfection, Luciferase, Reporter Assay, Activity Assay, Western Blot

Journal: Bioengineered

Article Title: CircRAB3IP upregulates twist family BHLH transcription factor (TWIST1) to promote osteosarcoma progression by sponging miR-580-3p.

doi: 10.1080/21655979.2021.1948487

Figure Lengend Snippet: Figure 5. miR-580-3p inhibitor or TWIST1 overexpression reversed effects of sh-circRAB3IP. MG63 cells were transfected with the control sh RNA (sh-NC), sh-circRAB3IP, sh-circRAB3IP+miR-580-3p inhibitor or sh-circRAB3IP+TWIST1 overexpression vector (sh- circRAB3IP+oeTWIST1). (a, b) Cell proliferation was detected by CCK-8 assay and colony formation assay. (c, d) Transwell assay was employed to detect cell migration and invasion of OS cells. *P < 0.05.

Article Snippet: Primary rabbit antihuman antibodies against TWIST1 (1:1000, cell signaling technology, #46702) and GAPDH (1:2000, Abcam, ab9485) were incubated overnight at 4°C.

Techniques: Over Expression, Transfection, Control, Plasmid Preparation, CCK-8 Assay, Colony Assay, Transwell Assay, Migration

Journal: Bioengineered

Article Title: Hsa_circ_0030586 promotes epithelial–mesenchymal transition in prostate cancer via PI3K-AKT signaling

doi: 10.1080/21655979.2021.2008217

Figure Lengend Snippet: Primers used in the study

Article Snippet: After blocking with 5% milk at room temperature for 2 h, primary antibodies targeting E-cadherin (dilution 1:5000; Proteintech, 20,874–1AP), Twist (dilution 1:1000; Proteintech, 25,465-1-AP), p-AKT (dilution 1:2000; CST, 4060s), AKT (dilution 1:2000; CST, 2920s), IKKα (dilution 1:10,000; Abcam, ab32041), PIK3CB (dilution 1:1000; Abcam, ab151549), and GAPDH (dilution 1:8000; Proteintech, 60,004-1-1 g) were incubated at 4°C overnight.

Techniques: Sequencing

Journal: Bioengineered

Article Title: Hsa_circ_0030586 promotes epithelial–mesenchymal transition in prostate cancer via PI3K-AKT signaling

doi: 10.1080/21655979.2021.2008217

Figure Lengend Snippet: Interfering with hsa_circ_0030586 suppressed the proliferation, migration, and invasion of PCa cells. (a) Transient interference fragments significantly reduced the expression of hsa_circ_0030586. (b) Transfection with hsa_circ_0030586 siRNA significantly inhibits cell viability at 72 h. (c) The EMT-related gene E-cadherin was significantly upregulated, and Twist was significantly downregulated in cells transfected with hsa_circ_0030586 siRNA. (d–e) Transfection with hsa_circ_0030586 siRNA significantly inhibited the migration and invasion of PC3 cells (scare bar: 100 μm). (f–g) Transfection with hsa_circ_0030586 siRNA significantly inhibited the wound healing capacity of PC3 cells (scare bar: 500 μm). *P < 0.05; ** P < 0.01

Article Snippet: After blocking with 5% milk at room temperature for 2 h, primary antibodies targeting E-cadherin (dilution 1:5000; Proteintech, 20,874–1AP), Twist (dilution 1:1000; Proteintech, 25,465-1-AP), p-AKT (dilution 1:2000; CST, 4060s), AKT (dilution 1:2000; CST, 2920s), IKKα (dilution 1:10,000; Abcam, ab32041), PIK3CB (dilution 1:1000; Abcam, ab151549), and GAPDH (dilution 1:8000; Proteintech, 60,004-1-1 g) were incubated at 4°C overnight.

Techniques: Migration, Expressing, Transfection

Journal: Bioengineered

Article Title: Hsa_circ_0030586 promotes epithelial–mesenchymal transition in prostate cancer via PI3K-AKT signaling

doi: 10.1080/21655979.2021.2008217

Figure Lengend Snippet: Hsa_circ_0030586 activates the PI3K-AKT signaling and promotes EMT. (a) The relative expression of five key molecules involved in the PI3K-AKT signaling pathway was detected by qRT-PCR. (b) The relative expression of three miRNAs targeting PIK3CB was detected by qRT-PCR. (c) The expression of E-cadherin, p-AKT, AKT, IKKα, PIK3CB, and Twist was tested via Western blot. GAPDH was used as an internal standard. (d) miR-145-3p expression was dectected in NC inhibitor and miR-145-3p inhibitor group; (e) The protein expression of E-cadherin, Twist, p-AKT, AKT, IKKα, and PIK3CB was tested via Western blot. GAPDH was used as an internal standard. *P < 0.05; ** P < 0.01

Article Snippet: After blocking with 5% milk at room temperature for 2 h, primary antibodies targeting E-cadherin (dilution 1:5000; Proteintech, 20,874–1AP), Twist (dilution 1:1000; Proteintech, 25,465-1-AP), p-AKT (dilution 1:2000; CST, 4060s), AKT (dilution 1:2000; CST, 2920s), IKKα (dilution 1:10,000; Abcam, ab32041), PIK3CB (dilution 1:1000; Abcam, ab151549), and GAPDH (dilution 1:8000; Proteintech, 60,004-1-1 g) were incubated at 4°C overnight.

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Bioengineered

Article Title: Hsa_circ_0030586 promotes epithelial–mesenchymal transition in prostate cancer via PI3K-AKT signaling

doi: 10.1080/21655979.2021.2008217

Figure Lengend Snippet: Interference with hsa_circ_0030586 inhibited tumorigenesis in nude mice. (a) Immunofluorescence staining shows the fluorescence rate of the cells. (b) Hsa_circ_0030586 expression in the shRNA and shNC groups was detected by qRT-PCR. (c) Nude mice and tumors injected with PC3-shRNA cells or PC3-shNC cells. (d, e) Tumor volumes (d) and weights (e) were calculated after nude mice were injected with PC3-shRNA cells or PC3-shNC cells. (f, g) p-AKT, AKT, IKKα, and PIK3CB protein expression was detected via Western blot. (h) HE and immunohistochemistry were used to detect the tumor morphology and expression of E-cadherin and Twist (scare bar: 100 μm). *p < 0.05; **p < 0.01

Article Snippet: After blocking with 5% milk at room temperature for 2 h, primary antibodies targeting E-cadherin (dilution 1:5000; Proteintech, 20,874–1AP), Twist (dilution 1:1000; Proteintech, 25,465-1-AP), p-AKT (dilution 1:2000; CST, 4060s), AKT (dilution 1:2000; CST, 2920s), IKKα (dilution 1:10,000; Abcam, ab32041), PIK3CB (dilution 1:1000; Abcam, ab151549), and GAPDH (dilution 1:8000; Proteintech, 60,004-1-1 g) were incubated at 4°C overnight.

Techniques: Immunofluorescence, Staining, Fluorescence, Expressing, shRNA, Quantitative RT-PCR, Injection, Western Blot, Immunohistochemistry

Journal: Cancer Cell International

Article Title: Identification of key genes as predictive biomarkers for osteosarcoma metastasis using translational bioinformatics

doi: 10.1186/s12935-021-02308-w

Figure Lengend Snippet: ARHGAP25 inhibited MG63 and U2OS cells’ growth, migration and invasion. a ARHGAP25 expression was determined in vector transfected MG63 and U2OS cells. b Overexpression of ARHGAP25 in MG63 and U2OS cells inhibited cell growth. c Representative images of MG63 and U2OS cells wound healing assay at 0 and 24 h after scratching. Bars indicate: 100 µm. d , e Quantification of the gap size in wound healing assay (n = 3), which showed that overexpression of ARHGAP25 suppressed wound healing. f Representative images of MG63 and U2OS cells stained with crystal violet in the transwell assay. Bars indicate: 100 µm. g Counts of cells invaded through the chambers (n = 3). h Representative images of colony formation assay to analyze cell proliferation in ARHGAP25 overexpressed OS cells. i Quantification of the colony number (n = 3). j Protein expression of Twist1, Vimentin, and E-cadherin was measured in OS cells with ARHGAP25 overexpression. k Quantification of protein expression using ImageJ. *Indicates P < 0.05, **indicates P < 0.01 and ***indicates P < 0.001. Picture c , f are magnified 100 times and h is not magnified

Article Snippet: Primary antibodies against TWIST1 (25465), anti-E-cadherin (20874), anti-vimentin (10366-1), and GAPDH (60004-1) were purchased from Proteintech (NJ, USA).

Techniques: Migration, Expressing, Plasmid Preparation, Transfection, Over Expression, Wound Healing Assay, Staining, Transwell Assay, Colony Assay

Journal: Journal of Cancer

Article Title: PI3K/Akt/HIF-1α signaling pathway mediates HPV-16 oncoprotein-induced expression of EMT-related transcription factors in non-small cell lung cancer cells

doi: 10.7150/jca.26112

Figure Lengend Snippet: HPV-16 oncoproteins enhanced Slug and Twist1 expression in NSCLC cells. A549 and NCI-H460 cells were transfected with plasmids harboring pEGFP-N1-HPV-16 E6 or E7, and the transfection with empty vector or mutant plasmids served as controls. (A) Western blot analysis was performed to detect Slug and Twist1 protein expression. (B) RT-qPCR was performed to determine Slug and Twist1 mRNA expression. All data are expressed as mean±SD of three independent experiments. Compared with empty vector and mutant controls, * P <0.05, ** P <0.01.

Article Snippet: Rabbit anti-human ZEB1, Snail1, Slug, and Twist1 primary antibodies and goat anti-rabbit HRP-conjugated secondary antibody were purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Expressing, Transfection, Plasmid Preparation, Mutagenesis, Western Blot, Quantitative RT-PCR

Journal: Journal of Cancer

Article Title: PI3K/Akt/HIF-1α signaling pathway mediates HPV-16 oncoprotein-induced expression of EMT-related transcription factors in non-small cell lung cancer cells

doi: 10.7150/jca.26112

Figure Lengend Snippet: Effect of HIF-1α knockdown on the protein expression of EMT-related transcription factors induced by HPV-16 oncoproteins in NSCLC cells. HPV-16 E6 (A and B)- or E7 (C and D)-transfected A549 and NCI-H460 cells were co-transfected with HIF-1α-siRNA or non-specific siRNA (NS-siRNA), followed by Western blot analysis for ZEB1, Snail1, Slug, and Twist1 protein expression. All results are representative of three independent experiments.

Article Snippet: Rabbit anti-human ZEB1, Snail1, Slug, and Twist1 primary antibodies and goat anti-rabbit HRP-conjugated secondary antibody were purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Knockdown, Expressing, Transfection, Western Blot

Journal: Journal of Cancer

Article Title: PI3K/Akt/HIF-1α signaling pathway mediates HPV-16 oncoprotein-induced expression of EMT-related transcription factors in non-small cell lung cancer cells

doi: 10.7150/jca.26112

Figure Lengend Snippet: Effect of HIF-1α knockdown on the mRNA expression of EMT-related transcription factors induced by HPV-16 oncoproteins in A549 NSCLC cells. HPV-16 E6 (A)- and E7(B)- transfected A549 cells were co-transfected with HIF-1α-siRNA or non-specific siRNA (NS-siRNA), followed by RT-qRCR analysis for ZEB1 , Snail1 , Slug, and Twist1 mRNA expression. All data are expressed as mean±SD of three independent experiments. * P <0.05, ** P <0.01.

Article Snippet: Rabbit anti-human ZEB1, Snail1, Slug, and Twist1 primary antibodies and goat anti-rabbit HRP-conjugated secondary antibody were purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Knockdown, Expressing, Transfection

Journal: Journal of Cancer

Article Title: PI3K/Akt/HIF-1α signaling pathway mediates HPV-16 oncoprotein-induced expression of EMT-related transcription factors in non-small cell lung cancer cells

doi: 10.7150/jca.26112

Figure Lengend Snippet: Effect of LY294002 on HPV-16 oncoprotein-induced Snail1, Slug, and Twist1 protein expression in NSCLC cells. HPV-16 E6 (A and B)- and E7- (C and D) transfected A549 and NCI-H460 cells were pretreated for 24 h with different concentrations of LY294002, a specific PI3K inhibitor, followed by Western blot analysis for Snail1, Slug, and Twist1 protein expression. All results are representative of three independent experiments.

Article Snippet: Rabbit anti-human ZEB1, Snail1, Slug, and Twist1 primary antibodies and goat anti-rabbit HRP-conjugated secondary antibody were purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Expressing, Transfection, Western Blot