Journal: Cell reports

Article Title: Breast cancer cell mesenchymal transition and metastasis directed by DAP5/eIF3d-mediated selective mRNA translation

doi: 10.1016/j.celrep.2023.112646

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-human/mouse Twist1 antibody , Abcam , Cat# ab50581/RRID:AB_883292.

Techniques: Recombinant, Staining, Protease Inhibitor, Virus, SYBR Green Assay, Bicinchoninic Acid Protein Assay, MTT Assay, Software, Imaging

Journal: Theranostics

Article Title: Vitamin D binding protein (VDBP) hijacks twist1 to inhibit vasculogenic mimicry in hepatocellular carcinoma

doi: 10.7150/thno.90322

Figure Lengend Snippet: GC interacts with Twist1 to inhibit Twist1-activated VE-cadherin transcription. (A) Silver staining of proteins acquired by FLAG-VDBP pull-down in cell lysis from MHCC-97H cells with Matrigel 3D culture. (B) Co-immunoprecipitation assays in MHCC-97H cells on Matrigel 3D culture with anti-VDBP followed by immunoblotting (IB) with antibodies against Twist1 and VDBP or with anti-Twist1 followed by IB with anti-Twist1 and VDBP. (C) Brightfield pictures (left) and fluorescent pictures of PLA assays with VDBP and Twist1 (right) in Twist1-negative cell (SNU-387) and Twist1-positive cell (SK-HEP-1) with Matrigel 3D culture. Scale bar, 40 µm for black and 5 µm for white. (D) Quantification graph of PLA signal per cell. n = 6, biological replicates. Statistics were calculated on biological replicates with two-tailed unpaired t -tests, *** P < 0.001. Error bars show mean with SD. (E) Visualization result of simulation docking between VDBP and Twist1. (F) Schematic diagram of full-length and truncated VDBP proteins. (G) Western blot analysis of Twist1 expression level in MHCC-97H cells with Matrigel 3D culture treated as indicated. (H) The transcriptional regulation of VE-cadherin by transfection of Twist1 and increasing VDBP plasmids detected with the dual-luciferase reporter assay. n = 6, biological replicates. Statistics were calculated on biological replicates with two-tailed unpaired t -tests, *** P < 0.001, compared with vector group, ## P < 0.01, ### P < 0.001, compared with the transfection of Twist1 alone group. Error bars show mean with SD. (I) Results of qPCR after Twist1 ChIP of CDH5 target gene in MHCC-97H cells with Matrigel 3D culture treated as indicated. n = 3, biological replicates. Statistics were calculated on biological replicates with two-tailed unpaired t -tests, ** P < 0.01, ns, not significant, compared with the control group. Error bars show mean with SD. (J) The levels of CDH5 mRNA were detected by RT-qPCR in MHCC-97H cells with Matrigel 3D culture treated as indicated. n = 3, biological replicates. Statistics were calculated on biological replicates with two-tailed unpaired t -tests, *** P < 0.001, ns, not significant, compared with the control group. Error bars show mean with SD. (K) The levels of VM related proteins were detected by western blot in MHCC-97H cells with Matrigel 3D culture treated as indicated.

Article Snippet: Rabbit polyclonal anti-Twist1 , , Proteintech , CAT#25465-1-AP , RRID: AB_2880093.

Techniques: Silver Staining, Lysis, Immunoprecipitation, Western Blot, Two Tailed Test, Expressing, Transfection, Luciferase, Reporter Assay, Plasmid Preparation, Quantitative RT-PCR

Journal: Theranostics

Article Title: Vitamin D binding protein (VDBP) hijacks twist1 to inhibit vasculogenic mimicry in hepatocellular carcinoma

doi: 10.7150/thno.90322

Figure Lengend Snippet: VDBP hijacks Twist1 to inhibit VM and suppresses the malignancy of HCC, supplementing VDBP can weaken the promoting effect of Twist1 on VM and malignant progression in HCC. (A and B) Representative plots of VM status of MHCC-97H cells at 0, 30, 120 and 240 min (A) and quantitative plots of the two measured parameters (B). Scale bar, 200 µm. (C) Fluorescent gelatin degradation and phalloidin/DAPI staining of MHCC-97H cells treated as indicated (top) and quantification of degradation area (bottom). Scale bar, 50 µm. n = 6, biological replicates. Statistics were calculated on biological replicates with two-tailed unpaired t -tests, *** P < 0.001, ns, not significant. Error bars show mean with SD. (D and E) Representative in vivo images (D) of MHCC-97H-luc-tumor-bearing-BALB/c-nude mice with indicated treatments and quantified values (E) are shown as relative luciferase activity (photons/area). n = 6, biological replicates. Statistics were calculated on biological replicates with two-tailed unpaired t -tests, * P < 0.05, ** P < 0.01, ns, not significant. Error bars show mean with SD. (F) Kaplan-Meier curves showing percentage of survival of MHCC-97H-luc-tumor-bearing-BALB/c-nude mice after indicated treatments. (G and H) Representative images (G) and quantification of pathological score (H) of IHC staining of VE-cadherin, E-cadherin, MMP2 and MMP9 in liver tissues of MHCC-97H-luc-tumor-bearing-BALB/c-nude mice. Scale bar, 40 µm. n = 6, biological replicates. Statistics were calculated on biological replicates with two-tailed unpaired t -tests, * P < 0.05, *** P < 0.001, compared with Control. ## P < 0.01, ### P < 0.001, ns, not significant. compared with oe Twist1. Error bars show mean with SD.

Article Snippet: Rabbit polyclonal anti-Twist1 , , Proteintech , CAT#25465-1-AP , RRID: AB_2880093.

Techniques: Staining, Two Tailed Test, In Vivo, Luciferase, Activity Assay, Immunohistochemistry

Journal: Theranostics

Article Title: Vitamin D binding protein (VDBP) hijacks twist1 to inhibit vasculogenic mimicry in hepatocellular carcinoma

doi: 10.7150/thno.90322

Figure Lengend Snippet: Materials

Article Snippet: Rabbit polyclonal anti-Twist1 , , Proteintech , CAT#25465-1-AP , RRID: AB_2880093.

Techniques: Luciferase, Reporter Gene Assay, Silver Staining, Binding Assay, Molecular Weight, In Situ, Membrane, Magnetic Beads, Transfection, Modification, H&E Stain, Staining, SYBR Green Assay, Bicinchoninic Acid Protein Assay, In Vivo, Plasmid Preparation, CRISPR, Knock-Out, Sequencing, Software, Imaging

Journal: iScience

Article Title: AKT2-mediated nuclear deformation leads to genome instability during epithelial-mesenchymal transition

doi: 10.1016/j.isci.2023.106992

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-Twist , Santa Cruz Biotechnology , Cat#sc-15393.

Techniques: Recombinant, Staining, Plasmid Preparation, Software, Imaging

Journal: iScience

Article Title: LRRC15 expression indicates high level of stemness regulated by TWIST1 in mesenchymal stem cells

doi: 10.1016/j.isci.2023.106946

Figure Lengend Snippet: Senescence-associated changes of human and mice MSCs (A) Morphological changes of the MSCs by continuous cultures. Senescent cells showed β-galactosidase expressions. Scale bar = 500 μm. (B) TWIST1/Twist1 expressions were suppressed in the senescent MSCs. Asterisk means significant difference detected at p < 0.05 (N = 3, biological replicates). (C) Gene expressions of stemness associated genes in the MSCs at each passage number. Asterisks mean significant difference were detected at p < 0.05 between control (N = 3, biological replicates).

Article Snippet: Rabbit Polyclonal anti - TWIST1 antibody , Cell Signaling , Cat# 46702; RRID: AB_2799308.

Techniques:

Journal: iScience

Article Title: LRRC15 expression indicates high level of stemness regulated by TWIST1 in mesenchymal stem cells

doi: 10.1016/j.isci.2023.106946

Figure Lengend Snippet: Suppression of TWIST1/Twist1 affected stem cell characteristics of the MSCs (A) Morphological changes of the MSCs by siTWIST1 treatment. Scale bar = 500 μm. (B) Relative cell numbers counted at 72 h after siRNA treatment in the mouse primary MSCs (mMSCs). Asterisks mean significant difference were detected at p < 0.05 between control (N = 3, biological replicates). (C) Relative cell numbers counted 72 h after siRNA treatment in human MSCs (hMSCs). Asterisks mean significant difference were detected at p < 0.05 between control (N = 3, biological replicates). (D) Western blot analysis of a cell cycle suppression and senescence marker P16INK4A in the siTWIST1-treated MSCs. (E) Heatmap comparisons of stemness-associated gene expressions between the scrambled RNA-transfected control and the siTWIST1/Twist1-treated MSCs both in human and mice. Numbers on the color scale are relative values to the no-treatment control as 1.0 and the values represent the average of three independent samples.

Article Snippet: Rabbit Polyclonal anti - TWIST1 antibody , Cell Signaling , Cat# 46702; RRID: AB_2799308.

Techniques: Western Blot, Marker, Transfection

Journal: iScience

Article Title: LRRC15 expression indicates high level of stemness regulated by TWIST1 in mesenchymal stem cells

doi: 10.1016/j.isci.2023.106946

Figure Lengend Snippet: Overexpression of TWIST1/Twist1 by a Sendai virus vector (SeV) in mouse and human MSCs (A) SeV resulted in high-efficient and stable gene introduction both in human and mice MSCs. Scale bar = 500 μm. (B) Efficiency of the SeV-TWIST1 transfection was observed by FACS analysis based on the GFP expression that was co-expressed with TWIST1. (C) TWIST1-overexpresing MSCs showed higher proliferation activity than the control cells transfected with SeV-GFP. Asterisks mean significant difference were detected at p < 0.05 (N = 3, biological replicates). (D) Western blot analysis of a proliferating cell marker PCNA in the TWIST1-overexpressing MSCs. (E) Gene expressions of the stemness-associated transcription factors and therapeutic cytokines in the SeV-GFP (control) and SeV-TWIST1-treated MSCs. Numbers on the color scale are relative values. Expression values used for the heatmap were the average obtained in three independent samples.

Article Snippet: Rabbit Polyclonal anti - TWIST1 antibody , Cell Signaling , Cat# 46702; RRID: AB_2799308.

Techniques: Over Expression, Plasmid Preparation, Transfection, Expressing, Activity Assay, Western Blot, Marker

Journal: iScience

Article Title: LRRC15 expression indicates high level of stemness regulated by TWIST1 in mesenchymal stem cells

doi: 10.1016/j.isci.2023.106946

Figure Lengend Snippet: LRRC15 correlated with the expression of TWIST1 in human and mouse MSCs (A) Heatmap analysis with two independent samples as replicate. Common trends were observed between same experimental group. (B) Gene ontology (GO) enrichment analysis of differentially upregulated genes in the TWIST1 overexpressing human bone marrow MSCs. (C) Scatterplot analysis of differentially expressed genes in the human bone marrow MSCs overexpressing TWIST1 or GFP. (D) Expression changes of LRRC15 gene under different gene expression levels of TWIST1 in human and mice MSCs. Y axis show relative expression level to non-treatment control cells. Asterisk means significant difference detected at p < 0.05 (N = 3, biological replicates). (E) Increased expression of LRRC15 protein in the TWIST1-overexpressing human MSCs.

Article Snippet: Rabbit Polyclonal anti - TWIST1 antibody , Cell Signaling , Cat# 46702; RRID: AB_2799308.

Techniques: Expressing

Journal: iScience

Article Title: LRRC15 expression indicates high level of stemness regulated by TWIST1 in mesenchymal stem cells

doi: 10.1016/j.isci.2023.106946

Figure Lengend Snippet: Characteristics of the LRRC15-positive cells in the mice bone marrow tissues (A) FACS analysis for the Lrrc15-expressing population in the CD11b/ − CD45 - /Ter119 - /7-AAD - fractions. (B) Comparison of Twist1 gene expression level between the Lrrc15 - and Lrrc15 + populations. Asterisk means significant difference were detected at p < 0.05 (N = 6 from different animals). (C) Colony formation properties of the FACS-sorted cells from CD11b/ − CD45 - /Ter119 - /7-AAD - /Lrrc15 - (upper) or CD11b − /CD45 - /Ter119 - /7-AAD - /Lrrc15 + (lower) fractions. (D) Flow cytometry analysis for the CD11b − /CD45 - /Ter119 - /7-AAD - /Lrrc15 + cells with the MSC markers in mice. Gray histograms show control cells treated with isotype antibodies, and pink histograms show samples treated with each antibody. (E) FACS sorting of the MSCs using anti-CD105 and anti-LRRC15 antibodies. (F) Comparing gene expressions of Twist1, stemness-associated transcription factors, and therapeutic cytokines in the CD11b − /CD45 - /Ter119 - /CD105 + /Lrrc15 - and CD11b − /CD45 - /Ter119 - /CD105 + /Lrrc15 + cells. Asterisks mean significant differences detected at p < 0.05 (N = 3, independent experiment). (G) Engrafted EGFP + cells were detected by FACS analysis. Numbers in the figures are mean values with S.D. from three independent animals per each group.

Article Snippet: Rabbit Polyclonal anti - TWIST1 antibody , Cell Signaling , Cat# 46702; RRID: AB_2799308.

Techniques: Expressing, Flow Cytometry

Journal: iScience

Article Title: LRRC15 expression indicates high level of stemness regulated by TWIST1 in mesenchymal stem cells

doi: 10.1016/j.isci.2023.106946

Figure Lengend Snippet: Characteristics of the LRRC15-positive cells in the human bone marrow tissues (A) FACS analysis showing the LRRC15 + population in the CD31 - /CD45 - /Ter119 - /7-AAD - and human MSC marker CD29 + fractions. (B) Comparing gene expressions of TWIST1 , other stemness-associated transcription factors, and therapeutic cytokines between the CD31 - /CD45 - /Ter119 - /CD29 + /LRRC15 - and the CD31 - /CD45 - /Ter119 - /CD29 + /LRRC15 + MSCs. (C) Relative gene expression levels of the signature genes in the CD31 - /CD45 - /Ter119 - /CD29 + /LRRC15 + cells collected from primary cultures of 5 different donors. Heatmap shows relative expression to the CD31 - /CD45 - /Ter119 - /CD29 + /LRRC15 - cells in the primary culture of same donors each. (D) Decrease of the LRRC15 + population by in vitro culture. (E) Ratios of the LRRC15 + MSCs at primary and passage 3. Asterisk means significant difference detected at p < 0.05 (N = 5 from different donors).

Article Snippet: Rabbit Polyclonal anti - TWIST1 antibody , Cell Signaling , Cat# 46702; RRID: AB_2799308.

Techniques: Marker, Expressing, In Vitro

Journal: iScience

Article Title: LRRC15 expression indicates high level of stemness regulated by TWIST1 in mesenchymal stem cells

doi: 10.1016/j.isci.2023.106946

Figure Lengend Snippet:

Article Snippet: Rabbit Polyclonal anti - TWIST1 antibody , Cell Signaling , Cat# 46702; RRID: AB_2799308.

Techniques: Software