Journal: Cell Death & Disease

Article Title: Targeting transglutaminase 2 mediated exostosin glycosyltransferase 1 signaling in liver cancer stem cells with acyclic retinoid

doi: 10.1038/s41419-023-05847-4

Figure Lengend Snippet: A Hierarchical clustering of fold change expression for the proteins measured by nLC-MS/MS in sorted EpCAM+ and EpCAM– JHH7 cells treated with vehicle control, 25 μM NC9, or 10 μM ACR for 16 h or shCtl and shTG2-transduced JHH7 cells. B Summary of the number of differentially expressed proteins with a threshold of more than 2-fold. C Comparison of downregulated proteins by NC9 and ACR in EpCAM+ JHH7 cells, downregulated proteins between shTG2 and shCtl JHH7 cells, and upregulated proteins between EpCAM+ and EpCAM– JHH7 cells. The three common proteins are highlighted. D Correlation between gene expression of TGM2 and EXT1 in a total of 25 HCC cell lines in the CCLE database. The data are presented as a robust multiarray average. E Protein expression of EXT1 in JHH7 cells treated with vehicle control, 25 μM NC9, or 25 μM ZDON for 16 h. F Gene expression of EXT1 in JHH7 cells treated with vehicle control, 25 μM NC9, or 25 μM ZDON for 4 h. G Gene expression of EXT1 and H cell proliferation of JHH7 cells transfected either with siCtl or siEXT1 for 24 h. Representative immunofluorescence staining for HS ( I ) in JHH7 cells treated with DMSO or 25 μM NC9 for 24 h and J in shCtl and shTG2-transduced JHH7 cells. Scale bar, 50 μm. K Immunofluorescence triple staining of DAPI (blue), TG2 (green), and HS (red) in EpCAM+ and EpCAM– JHH7 cells. Scale bar, 100 μm. L A schematic model of TGF-β1 activation-dependent regulatory network underlying the control of EXT1 gene expression by TG2 in HCC cells generated using “Upstream Regulator Analysis” and “Path Explore” functions in IPA platform. Gene expression of ( M ) EXT1 and 4 candidate upstream transcription regulators of EXT1 that CITED2 , KLF6 , HNF1B , and BHLHE40 and ( N ) downstream targets of TGF-β1 that SMAD2 , SMAD3 and SMAD4 in shCtl and shTG2-transduced JHH7 cells. ( O ) Gene expression of EXT1 and SMAD3 in JHH7 cells treated with DMSO or a TGF-β small molecule inhibitor SB431542 at 10 μM for 4 h. The data are presented as the mean ± SD; * P < 0.05, Student’s t -test.

Article Snippet: A pool of three target-specific siRNAs targeting human EXT1 (sc-36464, siEXT1) and control siRNA (sc-37007, siCtl) were purchased from Santa Cruz Biotechnology.

Techniques: Expressing, Tandem Mass Spectroscopy, Control, Comparison, Gene Expression, Transfection, Immunofluorescence, Staining, Activation Assay, Generated

Journal: Cell Death & Disease

Article Title: Targeting transglutaminase 2 mediated exostosin glycosyltransferase 1 signaling in liver cancer stem cells with acyclic retinoid

doi: 10.1038/s41419-023-05847-4

Figure Lengend Snippet: ACR directly bonded to and induced oligomer formation of TG2, which inhibited the transamidase activity of cytoplasmic TG2 in HCC cells. Loss-of-function of TG2 suppressed the expression of stemness-related genes, spheroid proliferation and selectively induced cell death of the EpCAM+ liver CSC subpopulation in HCC cells. Mechanistically, TG2 inhibition suppressed the gene and protein expression of the HS biosynthesis enzyme, EXT1, and HS biosynthesis in HCC cells. TG2 and HS were highly expressed and co-located in EpCAM+ liver CSCs, highlighting a potential role of TG2-mediated HSPG signaling in regulating cell proliferation of liver CSCs. In contrast, ACR at a high dose increased intracellular Ca2 + concentration along with clCasp3 + cells, which probably contributed to the enhanced transamidase activity of nuclear TG2, cross-linking of nuclear transcription factors such as Sp1, and apoptosis in HCC cells as previously reported [ , , ].

Article Snippet: A pool of three target-specific siRNAs targeting human EXT1 (sc-36464, siEXT1) and control siRNA (sc-37007, siCtl) were purchased from Santa Cruz Biotechnology.

Techniques: Activity Assay, Expressing, Inhibition, Concentration Assay

Journal: Journal of Animal Science

Article Title: Temperature Fluctuations Modulate Molecular Mechanisms in Skeletal Muscle and Influence Growth Potential in Beef Steers

doi: 10.1093/jas/skad343

Figure Lengend Snippet: TaqMan probes and primers used for RT-qPCR assays

Article Snippet: SCD , Bt04307476_m1.

Techniques: TaqMan Probe Assay, Sequencing

Journal: Journal of Animal Science

Article Title: Temperature Fluctuations Modulate Molecular Mechanisms in Skeletal Muscle and Influence Growth Potential in Beef Steers

doi: 10.1093/jas/skad343

Figure Lengend Snippet: mRNA expression and protein abundance levels of adipogenesis markers (PPARγ, C/EBPα, FAS, and SCD) in beef steers under temperature swing and transport conditions. ( A ) PPARγ , C/EBPα , FAS , and SCD mRNA levels in CON, TS, and TP groups. ( B ) Western blots (top) and quantification of protein levels (bottom) of PPARγ, C/EBPα, SCD, and FAS in CON, TS, and TP groups. Error bars indicate the standard error. The P -values are determined by one-way ANOVA (* P < 0.05, ** P < 0.01).

Article Snippet: SCD , Bt04307476_m1.

Techniques: Expressing, Quantitative Proteomics, Western Blot