Journal: Cell reports

Article Title: PAI-1 uncouples integrin-β1 from restrain by membrane-bound β-catenin to promote collagen fibril remodeling in obesity-related neoplasms

doi: 10.1016/j.celrep.2024.114527

Figure Lengend Snippet:

Article Snippet: Rabbit monoclonal anti-Pan-TEAD , Cell Signaling Technology , Cat# 13295S; RRID: AB_2687902.

Techniques: Recombinant, Membrane, Cell Culture, Reverse Transcription, SYBR Green Assay, Clinical Proteomics, Protein Extraction, Extraction, Bicinchoninic Acid Protein Assay, In Situ, Blocking Assay, Migration, shRNA, Control, Software, Pyromark Assay, Western Blot, Simple Western

Journal: Stem Cells Translational Medicine

Article Title: Extracellular vesicles conjugated with c(RGDyk) peptide targeting integrin αVβ3 repair optic nerve injury through YAP/TAZ and Smad2/3 signaling

doi: 10.1093/stcltm/szag006

Figure Lengend Snippet: Validation of target genes obtained via single-cell RNA sequencing in vivo/vitro models. Levels of interested proteins in (A) the retina and (B) optic nerve of ONC were presented. During recovery process by SC_EVs, protein levels involved in signal pathway were confirmed in (C) retina of ONC models and (D) R28 cells (* P < .05, ** P < .05 vs the age-matched sham; # P < .05, ## P < .01 vs EVs). At 9 h prior to SC_EVs treatment, R28 cells were exposed to 200 µM of CoCl 2 and 1 µM of TM2 TEAD inhibitor. Then, by (D) immunoblot and (E) immunofluorescence were performed for expression of proteins. The results are expressed as the mean ± standard error of the mean (SEM; scale bar: 10 μm). Statistical significance was determined using a nonparametric statistical test, followed by the Mann–Whitney U test (* P < .05, ** P < .01 vs control; # P < .05, ## P < .01vs CoCl 2 ; ++ P < .01 vs CoCl 2 +SC_EVs).

Article Snippet: Aliquots of 2 × 10 5 R28 cells were seeded onto glass coverslips and treated with 200 μM CoCl 2 and 1 μM of the TEA/TEF-domain (TEAD) transcription factor inhibitor TM2 (Tocris Bioscience, Bristol, UK) for 9 h. The hypoxia-damaged R28 cells were then incubated with EVs at a concentration of 24 μg/mL.

Techniques: Biomarker Discovery, Single Cell, RNA Sequencing, In Vivo, Western Blot, Immunofluorescence, Expressing, MANN-WHITNEY, Control

Journal: Acta Biochimica et Biophysica Sinica

Article Title: LncRNA AP000695.2 promotes glycolysis of lung adenocarcinoma via the miR-335-3p/TEAD1 axis

doi: 10.3724/abbs.2023227

Figure Lengend Snippet: AP000695.2 promotes TEAD1-mediated transcription of GLUT1 (A) Spearman correlation analysis in The Cancer Genome Atlas (TCGA) database showed that there is a positive correlation between AP000695.2 expression and TEAD1 mRNA expression (r = 0.2829). A significant positive correlation between the expression of TEAD1 mRNA and the mRNA expression of various glycolysis-related factors was observed. (B) Cellular experiments showed that the protein expression level of TEAD1 was increased after overexpression of AP000695.2, but decreased after the downregulation of AP000695.2. (C) In the GLUT1 Promoter, wt group, the expression of luciferase was significantly increased after overexpression of TEAD1. In the GLUT1 Promoter, mt group, the expression of luciferase was significantly increased after overexpression of TEAD1, and the expression level of luciferase in GLUT1 Promoter, wt group was significantly higher than that of GLUT1 Promoter, mt group after overexpression of TEAD1. MCS indicates negative control while TEAD1 indicates TEAD1 overexpression plasmid. (D) The expression of TEAD1 in TEAD1 overexpression (TEAD1) group was significantly higher than that in the NC group. (E) qPCR analysis showed that the mRNA expression level of GLUT1 in A549 and H1299 cell lines was increased after upregulating the expression of TEAD1. (F) Western blot analysis showed that the protein expression levels of GLUT1, HK2, PKM2 and LDHA in A549 and H1299 cell lines were increased after upregulating the expression of TEAD1. (G) Western blot analysis showed that the protein expression level of HIF-1α in A549 and H1299 cell lines was increased after upregulating the expression of TEAD1. * P<0.05, **P<0.01, ***P <0.001.

Article Snippet: TEAD1 overexpression plasmid (Origene, Rockville, USA) and si-TEAD1 (target sequence: 5′-GGATCCTCACAAGACGTCA-3′; RiboBio, Guangzhou, China) were used to regulate TEAD1 expression. miR-335-3p mimics, negative control (NC), miR-335-3p inhibitor and inhibitor NC (Synbio-tech) were used for the overexpression and knockdown of miR-335-3p.

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

Journal: Acta Biochimica et Biophysica Sinica

Article Title: LncRNA AP000695.2 promotes glycolysis of lung adenocarcinoma via the miR-335-3p/TEAD1 axis

doi: 10.3724/abbs.2023227

Figure Lengend Snippet: AP000695.2 promotes LUAD glycolysis, proliferation and migration via TEAD1 (A) The relative amount of FDG uptake of the L+sT group (transfected with AP000695.2 overexpression plasmid and TEAD1 siRNA) was lower than that of the L+s-NC (transfected with AP000695.2 overexpression plasmid and TEAD1 siRNA NC) group. The amount of FDG uptake in the s+T (transfected with AP000695.2 siRNA and TEAD1 overexpression plasmid) group was higher than that of the s+T-NC group (transfected with AP000695.2 siRNA and NC of TEAD1 overexpression plasmid). (B) The amount of lactic acid production in the L+sT group was lower than that in the L+s-NC group. The amount of lactic acid produced in the s+T group was higher than that in the s+T-NC group. (C) The level of glycolysis, glycolytic capacity and glycolytic reverse in the L+sT group were lower than those in the L+s-NC group. The level of glycolysis, glycolytic capacity and glycolytic reverse in the s+T group were higher than those of the s+T-NC group. (D) The cell proliferation ability of the L+sT group was lower than that of the L+s-NC group, and that of the s+T group was higher than that of the s+T-NC group on the 2nd and 3rd day after transfection. (E) The cell migration ability of the L+sT group was lower than that of the L+s-NC group, and that of the s+T group was higher than that of the s+T-NC group (magnification×200, scale bar: 100 μm). (F) The cell migration ability of the L+sT group was lower than that of the L+s-NC group, and that of the s+T group was higher than that of the s+T-NC group (magnification×100). **P<0.01, ***P <0.001.

Article Snippet: TEAD1 overexpression plasmid (Origene, Rockville, USA) and si-TEAD1 (target sequence: 5′-GGATCCTCACAAGACGTCA-3′; RiboBio, Guangzhou, China) were used to regulate TEAD1 expression. miR-335-3p mimics, negative control (NC), miR-335-3p inhibitor and inhibitor NC (Synbio-tech) were used for the overexpression and knockdown of miR-335-3p.

Techniques: Migration, Transfection, Over Expression, Plasmid Preparation, Produced

Journal: Acta Biochimica et Biophysica Sinica

Article Title: LncRNA AP000695.2 promotes glycolysis of lung adenocarcinoma via the miR-335-3p/TEAD1 axis

doi: 10.3724/abbs.2023227

Figure Lengend Snippet: miR-335-3p acts as a competing endogenous RNA participating in the regulation of TEAD1 and AP000695.2 (A) The binding site of TEAD1 and miR-335-3p and the binding site of miR-335-3p and AP000695.2. (B) qPCR experiment showed that the expression of miR-335-3p in A549 cells was decreased after upregulating AP000695.2 expression. The expression of miR-335-3p in H1299 cells was increased after AP000695.2 was downregulated. (C) The expression of miR-335-3p in A549 and H299 cells was increased after the transfection of miR-335-3p mimics. (D) Western blot analysis showed that the protein expression of TEAD1 was decreased after the transfection with miR-335-3p mimics, and increased after the transfection with miR-335-3p inhibitor, compared with the negative control group. (E) Dual-luciferase reporter assay indicated that miR-335-3p has binding sites with TEAD1 and AP000695.2. **P<0.01, ***P<0.001. NS, no significance.

Article Snippet: TEAD1 overexpression plasmid (Origene, Rockville, USA) and si-TEAD1 (target sequence: 5′-GGATCCTCACAAGACGTCA-3′; RiboBio, Guangzhou, China) were used to regulate TEAD1 expression. miR-335-3p mimics, negative control (NC), miR-335-3p inhibitor and inhibitor NC (Synbio-tech) were used for the overexpression and knockdown of miR-335-3p.

Techniques: Binding Assay, Expressing, Transfection, Western Blot, Negative Control, Luciferase, Reporter Assay

Journal: Acta Biochimica et Biophysica Sinica

Article Title: LncRNA AP000695.2 promotes glycolysis of lung adenocarcinoma via the miR-335-3p/TEAD1 axis

doi: 10.3724/abbs.2023227

Figure Lengend Snippet: Silencing of AP000695.2 inhibits tumor growth and aerobic glycolysis in vivo (A) The qPCR experiment showed that the expression of si-AP000695.2 group is significantly lower than that of negative control group. (B) The tumor volumes at different time points showed that from the 6th day after injection, compared with the si-NC group, the si-AP000695.2 significantly inhibited the growth of transplanted tumor. (C) The uptake of 18F-FDG by tumor was decreased after the expression of AP000695.2 was decreased. The picture on the left showed the micro-PET images. The right picture showed the quantitative analysis results of SUVmax. (D) Immunohistochemistry (IHC) staining showed that the expression levels of GLUT1, HK2, PKM2, LDHA and TEAD1 in si-AP000695.2 group were lower than those in si-NC group. (magnification × 400, scale bar: 100 μm). **P<0.01, ***P<0.001.

Article Snippet: TEAD1 overexpression plasmid (Origene, Rockville, USA) and si-TEAD1 (target sequence: 5′-GGATCCTCACAAGACGTCA-3′; RiboBio, Guangzhou, China) were used to regulate TEAD1 expression. miR-335-3p mimics, negative control (NC), miR-335-3p inhibitor and inhibitor NC (Synbio-tech) were used for the overexpression and knockdown of miR-335-3p.

Techniques: In Vivo, Expressing, Negative Control, Injection, Micro-PET, Immunohistochemistry

Journal: Acta Biochimica et Biophysica Sinica

Article Title: LncRNA AP000695.2 promotes glycolysis of lung adenocarcinoma via the miR-335-3p/TEAD1 axis

doi: 10.3724/abbs.2023227

Figure Lengend Snippet: Schematic diagram of the mechanisms of AP000695.2/miR-335-3p/TEAD1/GLUT1 axis in the glycolysis of LUAD AP000695.2 is involved in the mechanism of LUAD through functioning as a ceRNA to competitively sponge miR-335-3p, thereby regulating the expression of TEAD1. Then TEAD1 promotes glycolysis of lung adenocarcinoma by promoting GLUT1 transcription.

Article Snippet: TEAD1 overexpression plasmid (Origene, Rockville, USA) and si-TEAD1 (target sequence: 5′-GGATCCTCACAAGACGTCA-3′; RiboBio, Guangzhou, China) were used to regulate TEAD1 expression. miR-335-3p mimics, negative control (NC), miR-335-3p inhibitor and inhibitor NC (Synbio-tech) were used for the overexpression and knockdown of miR-335-3p.

Techniques: Expressing

Journal: Advanced Science

Article Title: CRISPR/Cas13 sgRNA‐Mediated RNA–RNA Interaction Mapping in Live Cells with APOBEC RNA Editing

doi: 10.1002/advs.202409004

Figure Lengend Snippet: sarID identifies NEAT1 ‐interacting transcripts in living cells. a) Read coverage at the AGO2 locus. C‐to‐U editing was indicated by the color. Brown and green indicate the abundances of C and U sites, respectively. b) Violin plot of C‐to‐U editing rates in the control and NEAT1 groups. n = 390 and 2341 for control and NEAT1 respectively; Wilcoxon test. c) Overlap of sites and genes identified in the control and NEAT1 groups. d) Metagene density plot of the distribution of C‐to‐U editing sites. e) Pathway enrichment analysis of interacting transcripts of NEAT1 . f) Scheme of the tRSA‐tagged pulldown assay. g) Real‐time RT‐PCR analysis of randomly selected interacting RNAs. GAPDH served as a negative control. The data were presented as mean ± SDs, n = 3 independent experiments, two‐tailed unpaired Student's t ‐test. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001. h,i) Real‐time RT‐PCR analysis (h) of the RNA levels of and western blot detection (i) of the protein levels of RIF1, XRN1, and TEAD1, when NEAT1 was overexpressed in 293T cells. j,k) Real‐time RT‐PCR analysis (j) of the RNA levels and western blot detection (k) of the protein levels of RIF1, XRN1, and TEAD1, when NEAT1 was knocked down in 293T cells. GAPDH was used as loading control in western blot.

Article Snippet: Primary antibodies used: GAPDH (Cell Signaling Technology Cat#2118), vinculin (Proteintech Cat#66305‐1‐Ig), NONO (Proteintech Cat#11058‐1‐AP), SFPQ (Proteintech Cat#15585‐1‐AP), Flag (Sigma, Cat#F1804), HNRNPK (Proteintech Cat#11426‐1‐AP), TEAD1 (Proteintech Cat#13283‐1‐AP), XRN1 (Bethyl Laboratories Cat# A300‐443A), RIF1 (Bethyl Laboratories Cat# A301‐102A), Lamin A/C (Cell Signaling Technology Cat# 2032).

Techniques: Control, Quantitative RT-PCR, Negative Control, Two Tailed Test, Western Blot