Journal: Cell Death & Disease

Article Title: GLI1 activation by non-classical pathway integrin α v β 3 /ERK1/2 maintains stem cell-like phenotype of multicellular aggregates in gastric cancer peritoneal metastasis

doi: 10.1038/s41419-019-1776-x

Figure Lengend Snippet: a Western blottingting showing upregulated Integrin α v β 3 , p-ERK1/2 and GLI1 in peritoneal MCAs, BGC823 MCAs and SGC7901 MCAs compared to scattered-free cancer cells in the control group. b Western blottingting showing downregulated GLI1 with Integrin α v β 3 inhibitor Cilengitide and upregulated GLI1 with co-stimulator ligand RGD compared to the blank control in SGC7901 MCAs and BGC823 MCAs. c Western blottingting showing the most pronounced effect of Integrin β 3 silencing in the shIntegrin β 3 #1 group. d Western blottingting showing downregulated GLI1 with Integrin α v β 3 co-stimulator ligand RGD plus ERK1/2 inhibitor PD-184161 and upregulated GLI1 with RGD alone compared to the blank control in SGC7901 MCAs and BGC823 MCAs. e Western blottingting showing slightly decreased GLI1 with Hedgehog/Smoothened pathway inhibitor Cyclopamine compared with the blank control group in SGC7901 MCAs and BGC823 MCAs. f , g Decreased luminescence of GLI1 in the group of Integrin α v β 3 co-simulator ligand RGD plus ERK1/2 inhibitor PD-184161 or PD-184161 alone compared to the blank control. Each bar in the figure represents the mean ± SEM of triplicates. * p < 0.05; ** p < 0.01

Article Snippet: The primary reagents used in this study were as follows: Integrin α v β 3 inhibitor Cilengitide (Cat#.S7077, Selleck) or Integrin α v β 3 co-stimulator ligand RGD (Arg-Gly-Asp) (Cat#.ab142698, Abcam) or ERK1/2 inhibitor PD-184161 (Cat#.ab143847, Abcam) or GLI1/2 inhibitor GANT61 (Cat.no.ab120904, Abcam) or Hedgehog/smoothen inhibitor Cyclopamine (Cat#.ab120392, Abcam).

Techniques: Western Blot, Control

Journal: Cell Death & Disease

Article Title: GLI1 activation by non-classical pathway integrin α v β 3 /ERK1/2 maintains stem cell-like phenotype of multicellular aggregates in gastric cancer peritoneal metastasis

doi: 10.1038/s41419-019-1776-x

Figure Lengend Snippet: a Decreased tumor spheres forming ability in the group of Integrin α v β 3 inhibitor Cilengitide or ERK1/2 inhibitor PD-184161 or GLI1 inhibitor GANT61 compared to the blank control in SGC7901 MCAs and BGC823 MCAs. Scale bar = 10μm. b Lower colony forming ability in the group of Integrin α v β 3 inhibitor Cilengitide or ERK1/2 inhibitor PD-184161 or GLI1 inhibitor GANT61 compared to the blank control in SGC7901 MCAs and BGC823 MCAs. c Western blottingting showing downregulated stemness-related genes CD44, ALDH1A1, Oct4, Bmi1 and Nanog in the group of Integrin α v β 3 inhibitor Cilengitide or ERK1/2 inhibitor PD-184161 or GLI1 inhibitor GANT61 compared to the blank control in SGC7901 MCAs and BGC823 MCAs. d , e Decreased volume and weight of xenograft tumors in the group of Integrinα v β 3 inhibitor Cilengitide or ERK1/2 inhibitor PD-184161 or GLI1 inhibitor GANT61 compared to the blank control in SGC7901 MCAs and BGC823 MCAs. Each bar in the figure represents the mean ± SEM of triplicates. * p < 0.05; ** p < 0.01

Article Snippet: The primary reagents used in this study were as follows: Integrin α v β 3 inhibitor Cilengitide (Cat#.S7077, Selleck) or Integrin α v β 3 co-stimulator ligand RGD (Arg-Gly-Asp) (Cat#.ab142698, Abcam) or ERK1/2 inhibitor PD-184161 (Cat#.ab143847, Abcam) or GLI1/2 inhibitor GANT61 (Cat.no.ab120904, Abcam) or Hedgehog/smoothen inhibitor Cyclopamine (Cat#.ab120392, Abcam).

Techniques: Control, Western Blot

Journal: Cell Death & Disease

Article Title: GLI1 activation by non-classical pathway integrin α v β 3 /ERK1/2 maintains stem cell-like phenotype of multicellular aggregates in gastric cancer peritoneal metastasis

doi: 10.1038/s41419-019-1776-x

Figure Lengend Snippet: a , b Higher abdominal cavity tumor nodules formed by overGLI1 and lower nodules formed by shIntegrin β 3 #1 and shGLI1#1 compared to the control group transduced SGC7901 MCAs and BGC823 MCAs. Arrows indicate intraperitoreal nodules. c Western blottingting showing higher levels of Integrin α v β 3 , p-ERK1/2, and GLI1 in fresh surgical specimens of GC peritoneal metastatic compared to paired primary tumors. Each bar in the figure represents the mean ± SEM of triplicates. * p < 0.05; ** p < 0.01

Article Snippet: The primary reagents used in this study were as follows: Integrin α v β 3 inhibitor Cilengitide (Cat#.S7077, Selleck) or Integrin α v β 3 co-stimulator ligand RGD (Arg-Gly-Asp) (Cat#.ab142698, Abcam) or ERK1/2 inhibitor PD-184161 (Cat#.ab143847, Abcam) or GLI1/2 inhibitor GANT61 (Cat.no.ab120904, Abcam) or Hedgehog/smoothen inhibitor Cyclopamine (Cat#.ab120392, Abcam).

Techniques: Control, Western Blot

Journal: Cell Death & Disease

Article Title: GLI1 activation by non-classical pathway integrin α v β 3 /ERK1/2 maintains stem cell-like phenotype of multicellular aggregates in gastric cancer peritoneal metastasis

doi: 10.1038/s41419-019-1776-x

Figure Lengend Snippet: Integrin α v β 3 mediates intercellular adhesion in MCAs of GC cells to form an independent functional unit and activates GLI1 through the non-classic ERK1/2 pathway and crosstalk with the classic Hedgehog pathway to maintain cancer stem cell-like phenotype

Article Snippet: The primary reagents used in this study were as follows: Integrin α v β 3 inhibitor Cilengitide (Cat#.S7077, Selleck) or Integrin α v β 3 co-stimulator ligand RGD (Arg-Gly-Asp) (Cat#.ab142698, Abcam) or ERK1/2 inhibitor PD-184161 (Cat#.ab143847, Abcam) or GLI1/2 inhibitor GANT61 (Cat.no.ab120904, Abcam) or Hedgehog/smoothen inhibitor Cyclopamine (Cat#.ab120392, Abcam).

Techniques: Functional Assay

Journal: Science Advances

Article Title: Dynamic adaptive coassembled sericin protein orchestrating stem cell development for nucleus pulposus regeneration

doi: 10.1126/sciadv.adx2768

Figure Lengend Snippet: ( A ) The composition and structure of the NP in IVD tissue exhibited spatial heterogeneity, which support NP cell development and physiological function through dynamic mechanical regulation and biochemical signaling. ( B ) Inspired by the partitioned structure of ECM, bioactive sericin was coassembled with BPAA-GFF fibrillar networks, driven by phase separation–induced molecular network rearrangement. The resulting dynamic adaptive network activated stem cell integrin ligands to modulate cell-matrix interactions and promoted chondrogenic differentiation via rapid stress relaxation and biochemical factor induction. ( C ) In rat and rabbit IVD degeneration models, KGN-incorporated coassembled hydrogel effectively promoted BMSCs for NP regeneration.

Article Snippet: To investigate the role of integrin β3 in mediating stem cell adhesion and cytoskeletal regulation, functional inhibition was performed using the αvβ3 integrin antagonist cilengitide (MedChemExpress, USA).

Techniques:

Journal: Science Advances

Article Title: Dynamic adaptive coassembled sericin protein orchestrating stem cell development for nucleus pulposus regeneration

doi: 10.1126/sciadv.adx2768

Figure Lengend Snippet: ( A ) 3D coculture of stem cells with the coassembled BS gel. ( B ) Time-dependent gene expression trend of stem cells regulated by coassembled BS protein gel. ( C ) Up-regulated genes and their associated biological events based on Gene Ontology (GO) analysis comparing BS versus B. ( D ) Flow relationships between activated molecular pathways and up-regulated genes based on Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis comparing BS versus B. JAK-STAT, Janus kinase–signal transducer and activator of transcription; MAPK, mitogen-activated protein kinase. ( E ) Gene interaction network analysis and Gantt chart analysis of temporal gene expression in three gene categories. ( F ) Gene expression heatmap analysis comparing BS versus B, n = 3 independent samples. ( G ) In vitro validation of gene expression by qPCR, n = 3 independent samples. ( H ) Local interaction residues in molecular docking. Color-coded representation shows BPAA-GFF (green), sericin protein (magenta), and the integrin αvβ3 complex (blue and orange). ( I ) Time-course analysis of stem cells cocultured with the hydrogel at 1, 2, and 4 weeks. ( J ) Integrin β3 expression analysis at 1 week, n = 3 independent samples. ( K ) Cytoskeletal staining and cell aspect ratio analysis at 1 week. ( L ) YAP staining and nuclear/cytoplasmic ratio analysis. ( M ) Coassembled BS gel activates integrin-mediated cell-matrix interactions, driving YAP nuclear translocation and downstream mechanotransduction signaling. Statistical analyses were performed with Student’s t tests (unpaired and two-tailed): * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: To investigate the role of integrin β3 in mediating stem cell adhesion and cytoskeletal regulation, functional inhibition was performed using the αvβ3 integrin antagonist cilengitide (MedChemExpress, USA).

Techniques: Gene Expression, In Vitro, Biomarker Discovery, Expressing, Staining, Translocation Assay, Two Tailed Test

Journal: Viruses

Article Title: Properties of Oligomeric Interaction of the Cytomegalovirus Core Nuclear Egress Complex (NEC) and Its Sensitivity to an NEC Inhibitory Small Molecule

doi: 10.3390/v13030462

Figure Lengend Snippet: Schematic illustration of the functional role and approaches for analysis of the human cytomegalovirus (HCMV) core nuclear egress complex (NEC). ( A ) The core of the HCMV-specific NEC is built by the pUL50–pUL53 complex (termed as groove and hook proteins, respectively) that recruits effector proteins of the multicomponent NEC, including the viral protein kinase pUL97, the bridging factor p32/gC1qR, emerin, protein kinase C (PKC)α, cyclin-dependent kinase 1 (CDK1) and the peptidyl-prolyl cis / trans isomerase Pin1. Moreover, hexameric arrangements of pUL50–pUL53 form a receptor lattice for the intranuclear docking of maturing progeny capsids, ultimately guiding them to inner nuclear membrane (INM) budding for perinuclear egress. At present, it is not known yet whether the HCMV-specific formation of a multicomponent NEC and a hexameric core NEC lattice are two processes taking place simultaneously or whether one of these depends on the other. Note, however, that this sophisticated functional interlocking of events, particularly the step of oligomeric pUL50–pUL53 assembly, has been considered as a sensitive target for NEC inhibitory small molecules. ( B ) A series of differentially tagged expression constructs were generated for the analysis of oligomeric interaction, including constructs for groove, hook and pUL50::pUL53 fusion proteins. ( C ) These constructs were used for the single transient expression of tagged versions of pUL50 and pUL53, to be analyzed in a newly established oligomeric NEC in vitro assembly assay. Oligomer formation could be demonstrated by coimmunoprecipitation and Western blot detection using the respective tag-specific antibodies.

Article Snippet: Monoclonal (mAb) and polyclonal (pAb) antibodies were used to detect the following cellular proteins: mouse mAb-emerin (H-12, Santa Cruz Biotech, Dallas, TX, USA), mouse mAb-p32 (anti-p32/gC1qR, Abcam, Cambridge, UK), mouse mAb-CDK1 (MA5-11472, Thermo Fisher), rabbit pAb-CDK1 (Sc-954, Santa Cruz Biotech), mouse mAb-PKCα (A-3, Santa Cruz Biotech), rabbit mAb-lamin A/C (EPR4100, Abcam) and mouse mAb-β-actin (AC-15, Sigma Aldrich).

Techniques: Functional Assay, Membrane, Expressing, Construct, Generated, In Vitro, Western Blot

Journal: Cell reports

Article Title: Cancer genes disfavoring T cell immunity identified via integrated systems approach

doi: 10.1016/j.celrep.2022.111153

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ABT199 (Selleckchem #S8048), ABT263 (Selleckchem #S1001), LCL161 (Selleckchem #S7009), Birinapant (Selleckchem #S7015), Fomepizole (Selleckchem #S1717), CHR2797 (Tocris #3595), XCT790 (Sigma Aldrich #X4753), Topiramate (Selleckchem #S1438), ACET (Tocris #2728), Almorexant (Selleckchem #S2160), SB334867 (R&D Systems, #1960), anti-IL1R1 (R&D Systems, #AF269), anti-IL9 (R&D Systems, #AF209), anti-ITGAV (Abcam, #ab16821), Cilengitide (Selleckchem, #S7077), Guanidine HCl (Sigma Aldrich, #G3272), Dalfampridine (Sigma Aldrich, #275875), BEZ235 (Selleckchem, #S1009), Apitolisib (Selleckchem, #S2696), S17092 (Sigma Aldrich, #SML0181), WNK463 (Selleckchem, #S8358).

Techniques: Functional Assay, Purification, Recombinant, Enzyme-linked Immunosorbent Assay, RNA Sequencing, Mutagenesis, CRISPR, Knock-Out, Construct, Software