Journal: Oncotarget

Article Title: FOXO4 expression is related to stem cell-like properties and resistance to treatment in diffuse large B-cell lymphoma

doi: 10.18632/oncotarget.13690

Figure Lengend Snippet: ( A ) Study scheme of gene expression analyses using doxorubicin (Dox) and phenylbutyrate (PB)-treated surviving cells: Four different cell lines (BJAB, Raji, Toledo, and Daudi) are treated with doxorubicin (300 nM) or phenylbutyrate (8 mM) for 48 h, and cDNA microarray analysis is done to identify differentially expressed target genes between treatment-surviving cells and parental control cells. ( B ) FOXO4 mRNA level is significantly higher in phenylbutyrate-treated surviving (PB) cells of BJAB, Raji and Daudi than control cells. ( C ) Vorinostat-treated surviving cells show higher mRNA level of FOXO4 than control (Con) cells of BJAB and Raji. ( D ) Primary lymphoma cells from three patients with refractory diffuse large B-cell lymphoma (DLBCL) show increased expression of FOXO4 in phenylbutyrate-treated surviving cells (PB) compared to the corresponding control cells. ( E ) The FOXO4 mRNA level is significantly higher in primary cells from four patients with refractory DLBCL than that of a patient with DLBCL who achieved complete response. Data represent means ± SD from three independent experiments.

Article Snippet: After endogenous peroxidase blocking for 5 min, tissues were incubated with primary polyclonal antibody against FOXO4 (1:1000; GTX50500; GeneTex, Irvine, CA, USA) for 15 min using a BOND-MAX autoimmunostainer (Leica Biosystems, Melbourne, Australia) for 15 min.

Techniques: Gene Expression, Microarray, Control, Expressing

Journal: Oncotarget

Article Title: FOXO4 expression is related to stem cell-like properties and resistance to treatment in diffuse large B-cell lymphoma

doi: 10.18632/oncotarget.13690

Figure Lengend Snippet: ( A, B ) Phenylbutyrate-treated surviving cells (BJAB-PB and Raji-PB) increase the expression of FOXO4 and its transcriptional targets (p21, p27, and SOD) compared to control cells. ( C, D ) BJAB-PB and Raji-PB cells show decreased expression of cyclin D1, CDK4, and cyclin A compared to control cells.

Article Snippet: After endogenous peroxidase blocking for 5 min, tissues were incubated with primary polyclonal antibody against FOXO4 (1:1000; GTX50500; GeneTex, Irvine, CA, USA) for 15 min using a BOND-MAX autoimmunostainer (Leica Biosystems, Melbourne, Australia) for 15 min.

Techniques: Expressing, Control

Journal: Oncotarget

Article Title: FOXO4 expression is related to stem cell-like properties and resistance to treatment in diffuse large B-cell lymphoma

doi: 10.18632/oncotarget.13690

Figure Lengend Snippet: ( A ) Soft agar colony formation in FOXO4-transfected or siFOXO4-transfected BJAB cells shows the amplification of FOXO4 increase colony forming ability and the knockdown of FOXO4 decrease colony formation in BJAB cell line. ( B ) Decrease in mRNA levels of Nanog, Oct-4 and Sox-2 in siFOXO4-transfected (siFOXO4) is noted compared to siControl-transfected (siCon) BJAB cells. ( C ) Phosphorylated AKT level is decreased in BJAB-PB cells with FOXO4 overexpression whereas siFOXO4-transfected BJAB-PB cells show the reverse of phosphorylated AKT. ( D ) The western blot shows the expression of FOXO4 protein in a BJAB clone (c2) overexpressing FOXO4. ( E ) Tumor sphere formation is observed from a BJAB clone (c2) overexpressing FOXO4. ( F ) Immunohistochemical staining for FOXO4 in tumor tissue of DLBCL (× 200). ( G ) Kaplan-Meier curves shows superior progression-free survival and overall survival of FOXO4-high group with diffuse large B cell lymphoma than FOXO4-low group. The P value is calculated using the log-rank test.

Article Snippet: After endogenous peroxidase blocking for 5 min, tissues were incubated with primary polyclonal antibody against FOXO4 (1:1000; GTX50500; GeneTex, Irvine, CA, USA) for 15 min using a BOND-MAX autoimmunostainer (Leica Biosystems, Melbourne, Australia) for 15 min.

Techniques: Transfection, Amplification, Knockdown, Over Expression, Western Blot, Expressing, Immunohistochemical staining, Staining

Journal: Oncotarget

Article Title: FOXO4 expression is related to stem cell-like properties and resistance to treatment in diffuse large B-cell lymphoma

doi: 10.18632/oncotarget.13690

Figure Lengend Snippet: Characteristics of patients

Article Snippet: After endogenous peroxidase blocking for 5 min, tissues were incubated with primary polyclonal antibody against FOXO4 (1:1000; GTX50500; GeneTex, Irvine, CA, USA) for 15 min using a BOND-MAX autoimmunostainer (Leica Biosystems, Melbourne, Australia) for 15 min.

Techniques:

Journal: Immunometabolism

Article Title: Metabolic Reprogramming during Microglia Activation

doi: 10.20900/immunometab20190002

Figure Lengend Snippet: Figure 10. Metabolic features of pro- and anti-inflammatory microglia. Pro-inflammatory microglia are characterized by increased glucose and glutamine metabolism (glycolysis, glucose oxidation through the TCA cycle and glutamine oxidation), while anti-inflammatory microglia upregulate fatty acid metabolism (mitochondrial fatty acid oxidation and fatty acid synthesis). Metabolic pathways upregulated in pro- inflammatory, M1 microglia or anti-inflammatory, M2 microglia are represented by red and blue color, respectively. Enzymes and starting substrates are marked in bold. ABCD-ATP-binding cassette transporters of subfamily D; ACOX1, acyl-CoA oxidase 1; CPT1, carnitine palmitoyltransferase 1; FASN, fatty acid synthase; G6PD, glucose-6-phosphate dehydrogenase; GLUD, glutamate dehydrogenase; GOT, glutamate-oxaloacetate transaminase; HK, hexokinase; LDH, lactate dehydrogenase; MCAD, medium-chain acyl-CoA dehydrogenase; MFP, multifunctional protein; PDH, pyruvate dehydrogenase; PDK, pyruvate dehydrogenase kinase; PFKFB, phosphofructokinase/fructose biphosphatase; TAG, triacylglycerides; TCA, tricarboxylic acid cycle; VLCFA, very long chain fatty acids.

Article Snippet: The following primary antibodies were used, ACOX1 (against 51 kDa subunit; 1/5000 [42]), MFP2 (1/400, ProteinTech, Manchester, UK) and vinculin (1/2000, Sigma Aldrich, St. Louis, MO, USA) together with HRP-conjugated secondary antibodies (1/5000, Agilent, Santa Clara, CA, D ow nloaded from http://journals.lw w .com /im m unom etabolism by B hD M f5eP H K av1zE oum 1tQ fN 4a+ kJLhE Z gbsIH o4X M i0hC yw C X 1A W nY Q p/IlQ rH D 3i3D 0O dR yi7T vS F l4C f3V C 1y0abggQ Z X dgG j2M w lZ LeI= on 01/18/2024 Immunometabolism.

Techniques: Binding Assay

Journal: Molecular Medicine Reports

Article Title: Overexpression of long noncoding RNA CUDR promotes hepatic differentiation of human umbilical cord mesenchymal stem cells

doi: 10.3892/mmr.2019.10897

Figure Lengend Snippet: Sequences of reverse transcription-quantitative PCR primers.

Article Snippet: Primary antibodies against hepatocyte nuclear factor 4α (HNF4α, sc-374299, Santa Cruz Biotechnology, Inc.), hepatocyte nuclear factor 3β (HNF3β, sc-374376, Santa Cruz Biotechnology, Inc.), hepatocyte nuclear factor 6 (HNF6, sc-365318, Santa Cruz Biotechnology, Inc.), enhanced binding protein α (CEBPα, sc-133239, Santa Cruz Biotechnology, Inc.), β-catenin (sc-47724, Santa Cruz Biotechnology, Inc.) and GAPDH (all at 1:2,000 dilution) were incubated overnight at 4°C and washed with TBS containing 0.1% Tween-20.

Techniques: Sequencing

Journal: Molecular Medicine Reports

Article Title: Overexpression of long noncoding RNA CUDR promotes hepatic differentiation of human umbilical cord mesenchymal stem cells

doi: 10.3892/mmr.2019.10897

Figure Lengend Snippet: Hepatic differentiation of HuMSCs. (A) HuMSCs were identified by osteogenic and chondrogenic differentiation. (B) Morphological changes observed as HuMSCs differentiated into hepatocytes with the addition of cytokines. (C) mRNA levels of ALB, AFP, CYP3A4 and HNF4α after differentiation were analyzed via RT-qPCR. (D) ALB, AFP, CYP3A4 protein levels after differentiation were analyzed by immunocytochemistry. (E) Expression of CUDR was analyzed via RT-qPCR. All experiments were performed three times independently. *P<0.05 vs. Day 0 group. All experiments were performed three independent times. HuMSCs, human umbilical cord mesenchymal stem cells; ALB, albumin; AFB, α fetoprotein; CYP3A4, cytochrome P450 3A4; HNF4α, hepatocyte nuclear factor 4α; RT-qPCR, reverse transcription-quantitative PCR; CUDR, cancer upregulated drug resistant.

Article Snippet: Primary antibodies against hepatocyte nuclear factor 4α (HNF4α, sc-374299, Santa Cruz Biotechnology, Inc.), hepatocyte nuclear factor 3β (HNF3β, sc-374376, Santa Cruz Biotechnology, Inc.), hepatocyte nuclear factor 6 (HNF6, sc-365318, Santa Cruz Biotechnology, Inc.), enhanced binding protein α (CEBPα, sc-133239, Santa Cruz Biotechnology, Inc.), β-catenin (sc-47724, Santa Cruz Biotechnology, Inc.) and GAPDH (all at 1:2,000 dilution) were incubated overnight at 4°C and washed with TBS containing 0.1% Tween-20.

Techniques: Quantitative RT-PCR, Immunocytochemistry, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Overexpression of long noncoding RNA CUDR promotes hepatic differentiation of human umbilical cord mesenchymal stem cells

doi: 10.3892/mmr.2019.10897

Figure Lengend Snippet: CUDR promotes hepatic differentiation by regulating liver-enriched factors. (A) mRNA of HNF4α, HNF3β, HNF6 and CEBPα were analyzed by reverse transcription-quantitative PCR after 14 and 28 days of differentiation. (B) Protein levels of HNF4α, HNF3β, HNF6 and CEBPα were analyzed by western blot analysis after 14 and 28 days of differentiation. *P<0.05 vs. control group. All experiments were performed three independent times. HNF4α, hepatocyte nuclear factor 4α; CUDR, cancer upregulated drug resistant; HNF3β, hepatocyte nuclear factor 3β; HNF6, hepatocyte nuclear factor 6; CEBPα, enhanced binding protein α.

Article Snippet: Primary antibodies against hepatocyte nuclear factor 4α (HNF4α, sc-374299, Santa Cruz Biotechnology, Inc.), hepatocyte nuclear factor 3β (HNF3β, sc-374376, Santa Cruz Biotechnology, Inc.), hepatocyte nuclear factor 6 (HNF6, sc-365318, Santa Cruz Biotechnology, Inc.), enhanced binding protein α (CEBPα, sc-133239, Santa Cruz Biotechnology, Inc.), β-catenin (sc-47724, Santa Cruz Biotechnology, Inc.) and GAPDH (all at 1:2,000 dilution) were incubated overnight at 4°C and washed with TBS containing 0.1% Tween-20.

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Control, Binding Assay

Journal: Molecular Medicine Reports

Article Title: Overexpression of long noncoding RNA CUDR promotes hepatic differentiation of human umbilical cord mesenchymal stem cells

doi: 10.3892/mmr.2019.10897

Figure Lengend Snippet: CUDR inhibits Wnt signaling activity by decreasing β-catenin cyto-nuclear translocation. (A) Immunocytochemistry staining for the localization of β-catenin in hepatocyte-like cells after differentiation when transfected with control or CUDR. (B) TOP/FOP assay showing the decreased activity of Wnt signaling in the hepatocyte-like cells with overexpression of CUDR. (C) RT-qPCR analysis showing CUDR transduction downregulated the mRNA expression of β-catenin. (D) Result of western blot analysis was in accordance with RT-qPCR. *P<0.05 vs. control group. All experiments were performed three independent times. CUDR, cancer upregulated drug resistant; RT-qPCR, reverse transcription-quantitative PCR.

Article Snippet: Primary antibodies against hepatocyte nuclear factor 4α (HNF4α, sc-374299, Santa Cruz Biotechnology, Inc.), hepatocyte nuclear factor 3β (HNF3β, sc-374376, Santa Cruz Biotechnology, Inc.), hepatocyte nuclear factor 6 (HNF6, sc-365318, Santa Cruz Biotechnology, Inc.), enhanced binding protein α (CEBPα, sc-133239, Santa Cruz Biotechnology, Inc.), β-catenin (sc-47724, Santa Cruz Biotechnology, Inc.) and GAPDH (all at 1:2,000 dilution) were incubated overnight at 4°C and washed with TBS containing 0.1% Tween-20.

Techniques: Activity Assay, Translocation Assay, Immunocytochemistry, Staining, Transfection, Control, Over Expression, Quantitative RT-PCR, Transduction, Expressing, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction