Journal: Cellular and molecular gastroenterology and hepatology

Article Title: Additive Effect of CD73 Inhibitor in Colorectal Cancer Treatment With CDK4/6 Inhibitor Through Regulation of PD-L1.

doi: 10.1016/j.jcmgh.2022.07.005

Figure Lengend Snippet: Figure 8. Negative corre- lation between ecto- enzymes and CCND1 expression in CRC. (A) IHC staining of human normal colon (n ¼ 40) and colon adenocarcinoma (n ¼ 64) specimens in a tissue microarray for CCND1 and CD163 (a macrophage marker). (B) Relative intensities of CCND1 staining in the lamina propria of a normal and cancerous colon. **P < .01 (unpaired t test). (C) Correlation analysis of NT5E and CCND1 expres- sion in the TCGA colorectal adenocarcinoma dataset. ***P < .001 (Student t test).

Article Snippet: For the fluorescence-activated cell sorting analysis, cells were stained with APC,-anti human CD163 (clone: GHI/61), PD-anti human CTLA4 (clone: L3D10), Alexa 647-anti human IDO1 (clone: 2E2/IDO1), PE-anti human DR4 (clone: DJR1), FITC-anti human CD47 (clone: REA220, FITC-anti human MICA&B (clone: 6D4), PE-anti human PD-L1 (clone: MIH2), PD-anti human CD69 (clone: FNM50), FITCanti human CD2 (clone: RPA-2.10), FITC-anti human CD20 (clone: 2H7), APC-anti mouse F4/80 (clone: BM8), Brilliant Violet 421TM-anti mouse CD11b (clone: M1/70), Brilliant Violet 570TM-anti mouse CD45 (clone: 104), PE-anti-human CD45 (clone: HI30), monoclonal antibodies (Biolegend; San Diego, CA, USA), FITC-anti-human CD40 (clone: REA733), PE-anti human CD80 (clone: 2D10), PE/vio770-anti human CD206 (clone: DCN228), PE-anti human CD62E (clone: REA280), PE-anti human CD192 (clone: REA264), APC-anti human I-CAM (clone: REA266), APC-anti human HLA-DR, DP, DQ (clone: REA332), APC-anti human CCL2 (clone: REA485), APC-anti human CD14 (clone: HI30), Vioblue-anti human CD31 (clone: TUK4), FTIC-anti-human CD86 (clone: FM95), PE-anti human CD73 (clone: AD2), PE-anti mouse PD-L1 (clone: 60533), PE-anti mouse CD39, and PE-anti human CD39 (clone: REA739) monoclonal antibodies (Miltenyi Biotec; Bergisch Gladbach, Germany), and PE/CF594anti human CD3 (clone: UCHT1), PE-anti human CD44 (clone: G44-26), PE-anti human CD183 (clone: 150503), and PD-1 (clone: REA739) monoclonal antibodies (BD Biosciences; San Jose, CA, USA) following the manufacturer’s protocol.

Techniques: Expressing, Immunohistochemistry, Microarray, Marker, Staining

Journal: OncoTargets and therapy

Article Title: Overexpression of UHRF2 in intrahepatic cholangiocarcinoma and its clinical significance

doi: 10.2147/OTT.S149361

Figure Lengend Snippet: The expression of UHRF2 in human cancers and ICC patients. Notes: ( A ) Microarray data analysis of UHRF2 mRNA in gastric cancer, colon cancer, and hepatocellular carcinoma from the publicly available Oncomine database. ( B ) UHRF2 protein expression in randomly selected ICC tissues and their matched peritumor tissues by Western blot. ( C ) Relative expression of UHRF2 among tumor and peritumor tissues using qRT-PCR. ( D ) Positive UHRF2 was observed primarily in the cytoplasm. Representative UHRF2 and H

Article Snippet: Rabbit anti-human UHRF2 monoclonal antibody (1:1,000; Wuhan Boster Biological Technology, Ltd., Wuhan, People’s Republic of China) and rabbit anti-human E-cadherin antibody (1:100; Cell Signaling Technology, Danvers, MA, USA) were utilized to detect the levels of UHRF2 and E-cadherin.

Techniques: Expressing, Microarray, Western Blot, Quantitative RT-PCR, Staining, Real-time Polymerase Chain Reaction, Ubiquitin Proteomics, Immunohistochemistry

Journal: OncoTargets and therapy

Article Title: Overexpression of UHRF2 in intrahepatic cholangiocarcinoma and its clinical significance

doi: 10.2147/OTT.S149361

Figure Lengend Snippet: UHRF2 expression resulted in ICC cell proliferation, invasion, migration, and antiapoptosis. Notes: ( A ) UHRF2 expression was interfered by siRNA and confirmed by Western blot and qRT-PCR. GAPDH was used as internal control. ( B ) Cell counting kit-8 assay was used to assess the ability of cell proliferation. ( C ) Transwell assay was used to show the invasion of ICC cells with different UHFR2 expression in 48 hours. Scale bar =100 μm. ( D ) Wound healing assays showed that inhibition of UHRF2 decreased wound healing compared with control cells. Scale bar =100 μm. ( E ) FCM results indicated that anti-UHRF2 caused acceleration of cell apoptosis. The results are mean ± SD of triplicated independent experiments. * P <;0.05, ** P <;0.01, *** P <;0.001. Abbreviations: FCM, flow cytometry; FITC, fluorescein isothiocyanate; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; ICC, intrahepatic cholangiocarcinoma; qRT-PCR, quantitative real-time polymerase chain reaction; UHRF2, ubiquitin-like with PHD and ring finger domains 2.

Article Snippet: Rabbit anti-human UHRF2 monoclonal antibody (1:1,000; Wuhan Boster Biological Technology, Ltd., Wuhan, People’s Republic of China) and rabbit anti-human E-cadherin antibody (1:100; Cell Signaling Technology, Danvers, MA, USA) were utilized to detect the levels of UHRF2 and E-cadherin.

Techniques: Expressing, Migration, Western Blot, Quantitative RT-PCR, Control, Cell Counting, Transwell Assay, Inhibition, Flow Cytometry, Real-time Polymerase Chain Reaction, Ubiquitin Proteomics

Journal: OncoTargets and therapy

Article Title: Overexpression of UHRF2 in intrahepatic cholangiocarcinoma and its clinical significance

doi: 10.2147/OTT.S149361

Figure Lengend Snippet: UHRF2 was negatively associated with E-cadherin expression in ICC. Notes: ( A ) High expression of UHRF2 and low expression of E-cadherin, and low expression of UHRF2 and high expression of E-cadherin are shown. Scale bar: 100, 200 μm. ( B ) The relationship between UHRF2 and E-cadherin in ICC patients; the protein expression of UHRF2 was significantly negatively correlated with E-cadherin expression. ( C ) UHRF2 and E-cadherin protein were detected by Western blot analysis. ( D ) Representative immunofluorescent images of UHRF2 and E-cadherin in ICC cell lines are shown. Scale bars =100 μm. * P <;0.05, ** P <;0.01, *** P <;0.001. Abbreviations: H

Article Snippet: Rabbit anti-human UHRF2 monoclonal antibody (1:1,000; Wuhan Boster Biological Technology, Ltd., Wuhan, People’s Republic of China) and rabbit anti-human E-cadherin antibody (1:100; Cell Signaling Technology, Danvers, MA, USA) were utilized to detect the levels of UHRF2 and E-cadherin.

Techniques: Expressing, Western Blot, Ubiquitin Proteomics

Journal: OncoTargets and therapy

Article Title: Overexpression of UHRF2 in intrahepatic cholangiocarcinoma and its clinical significance

doi: 10.2147/OTT.S149361

Figure Lengend Snippet: Prognostic significance was assessed by log-rank tests and Kaplan–Meier analysis. Notes: ( A – D ) Representative graphs of H

Article Snippet: Rabbit anti-human UHRF2 monoclonal antibody (1:1,000; Wuhan Boster Biological Technology, Ltd., Wuhan, People’s Republic of China) and rabbit anti-human E-cadherin antibody (1:100; Cell Signaling Technology, Danvers, MA, USA) were utilized to detect the levels of UHRF2 and E-cadherin.

Techniques: Immunohistochemical staining, Staining, Expressing, Ubiquitin Proteomics

Journal: OncoTargets and therapy

Article Title: Overexpression of UHRF2 in intrahepatic cholangiocarcinoma and its clinical significance

doi: 10.2147/OTT.S149361

Figure Lengend Snippet: Correlations between UHRF2 with clinicopathologic features in 139 ICC patients

Article Snippet: Rabbit anti-human UHRF2 monoclonal antibody (1:1,000; Wuhan Boster Biological Technology, Ltd., Wuhan, People’s Republic of China) and rabbit anti-human E-cadherin antibody (1:100; Cell Signaling Technology, Danvers, MA, USA) were utilized to detect the levels of UHRF2 and E-cadherin.

Techniques:

Journal: OncoTargets and therapy

Article Title: Overexpression of UHRF2 in intrahepatic cholangiocarcinoma and its clinical significance

doi: 10.2147/OTT.S149361

Figure Lengend Snippet: Univariate and multivariate analyses of factors associated with recurrence and survival

Article Snippet: Rabbit anti-human UHRF2 monoclonal antibody (1:1,000; Wuhan Boster Biological Technology, Ltd., Wuhan, People’s Republic of China) and rabbit anti-human E-cadherin antibody (1:100; Cell Signaling Technology, Danvers, MA, USA) were utilized to detect the levels of UHRF2 and E-cadherin.

Techniques:

Journal: European journal of cell biology

Article Title: The transcriptional profile of adipose-derived stromal cells (ASC) mirrors the whitening of adipose tissue with age.

doi: 10.1016/j.ejcb.2022.151206

Figure Lengend Snippet: Fig. 4. Adipose-derived stromal cells (ASC) resident in white adipose tissue (WAT) switch from a brown- to white-like precursor transcriptional signature with age. Differential expression analyses of brown adipose tissue (BAT)- and WAT-derived precursor cells in the GEO-GSE7032 dataset identified multiple genes that were significantly (t-test p < 0.05) overexpressed or under-expressed in undifferentiated/early differentiated BAT precursor cells (BAT precursor signature) compared to undifferentiated/early differentiated WAT precursor cells (WAT precursor signature) (A). Gene set enrichment analysis (GSEA) revealed that the BAT precursor signature was enriched in ASCs from the inguinal WAT of 1-month-old mice, while the WAT precursor signature was enriched in ASCs from the inguinal WAT of 12- month-old mice (B). The precursor signatures of WAT and BAT were refined to include only the genes that displayed significant individual core enrichment in WAT- resident ASCs from either 1-month-old mice (UP in BAT/1mo signature) or 12-month-old mice (UP in WAT/12mo signature) according the GSEA metrics (C). UP in BAT/1mo and UP in WAT/12mo signatures were compared with adipose tissue cold-induced transcriptional changes reported by the studies of Roh et al. (GSE108077) and Zhu et al. (GSE84860) (D). Analysis with METASCAPE algorithm (Zhou et al., 2019) (https://metascape.org/gp/index.html#/main/step1) revealed that the UP in BAT/1mo signatures were mainly enriched in genes related to mitochondrial biogenesis and function (E). Knock-down of Interferon Regulatory Factor 7 (Irf7) did not change the expression profile of UP in BAT/1mo and UP in WAT/12mo signatures in 12mo ASC (F). TRRUST(Han et al., 2018) analysis through METASCAPE revealed two clusters of transcription factors the targets of which were differently enriched in UP in WAT/12mo or BAT/1mo signatures. Analysis with the STRING(Szklarczyk et al., 2017) database (https://string-db.org/) identified multiple interactions previously reported between these transcription factors (G). Luciferase assay of cells engineered with Luciferase gene under the control of either E2F1 or NFkB consensus binding sequence revealed a lower transcriptional activation by E2F1 and a higher transcriptional activity of NFkB in 12mo ASC compared to 1mo ASC (H).

Article Snippet: To assess the expression of miR200a,429,200b family cluster, cells were transiently transfected with pGL3–1574/+ 120, which encodes Firefly Luciferase downstream a segment of the human gene, encompassing − 1574 to + 120 relative to the putative transcription start site (TSS) (Bracken et al., 2008) of miR 200a, 200b, 429 family cluster. pGL3–1574/+ 120 was a gift from Greg Goodall (Addgene plasmid # I. Scambi et al. European Journal of Cell Biology 101 (2022) 151206 35539; http://n2t.net/addgene:35539; RRID:Addgene_35539).

Techniques: Derivative Assay, Quantitative Proteomics, Knockdown, Expressing, Luciferase, Control, Binding Assay, Sequencing, Activation Assay, Activity Assay

Journal: European journal of cell biology

Article Title: The transcriptional profile of adipose-derived stromal cells (ASC) mirrors the whitening of adipose tissue with age.

doi: 10.1016/j.ejcb.2022.151206

Figure Lengend Snippet: Fig. 5. Aging of the adipose-derived stromal cells (ASC) resident in white adipose tissue (WAT) brings about an overall down-regulation of miRNAs. The microarray profile of miRNA expression in ASCs taken from the inguinal WAT of mice aged 1, 3, 6, 12 or 24 months showed an overall decline of miRNA with age. The list of miRNAs that were downregulated with age included many of the miRNAs the depletion of which was previously shown to be implicated in the brown-to-white switch of adipocyte precursors (Mori et al., 2014) (A). Previously validated targets of these miRNAs according to miRtarBase (Targets miRNA down) were overrepresented in the transcriptome of WAT-resident ASCs from 12-month-old mice compared to 1-month-old mice (B). Luciferase assay of ASC engineered with Firefly luciferase gene downstream a segment of the human gene, encompassing − 1574 to + 120 relative to the putative transcription start site (TSS) (Bracken et al., 2008) of miR200a, 200b,429 family cluster revealed a higher transcriptional activity of this cluster in 1mo ASC compared to 12mo ASC (C). In silico anaysis of transcription factor binding site enrichment through TransmiR 2.0 algorithm (Tong et al., 2019) (http://www.cuilab.cn/transmir) revealed that miRNA significantly anticorrated with age (Correlation index, Ci<−0.7, p < 0.05) were targets of a limited number of transcription factors, including NFkB, RELA and E2F1 (D).

Article Snippet: To assess the expression of miR200a,429,200b family cluster, cells were transiently transfected with pGL3–1574/+ 120, which encodes Firefly Luciferase downstream a segment of the human gene, encompassing − 1574 to + 120 relative to the putative transcription start site (TSS) (Bracken et al., 2008) of miR 200a, 200b, 429 family cluster. pGL3–1574/+ 120 was a gift from Greg Goodall (Addgene plasmid # I. Scambi et al. European Journal of Cell Biology 101 (2022) 151206 35539; http://n2t.net/addgene:35539; RRID:Addgene_35539).

Techniques: Derivative Assay, Microarray, Expressing, Luciferase, Activity Assay, In Silico, Binding Assay

Journal: Cell

Article Title: Mapping Systemic Inflammation and Antibody Responses in Multisystem Inflammatory Syndrome in Children (MIS-C)

doi: 10.1016/j.cell.2020.09.034

Figure Lengend Snippet:

Article Snippet: anti-GranzymeB 111Cd-conjugated Clone REA226 , Miltenyi , Cat No.130-108-055; RRID: AB_2659980.

Techniques: Virus, Recombinant, Saline, Blocking Assay, Staining, Microarray, Software, Immunoprecipitation

Journal: Cell reports

Article Title: TIFAB Regulates USP15-Mediated p53 Signaling during Stressed and Malignant Hematopoiesis

doi: 10.1016/j.celrep.2020.01.093

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Ubiquitin-Histone 2B (Lys 120) antibody , Cell Signaling , Cat# 5546P.

Techniques: Ubiquitin Proteomics, Virus, Plasmid Preparation, Recombinant, Silver Staining, Transfection, Purification, Lysis, Microarray, Negative Control, Software