Journal: Nutrients

Article Title: Integrated Proteomics and Metabolomics Reveal the Direct Hepatic Protection of Propionate Against Alcoholic Liver Disease via the RGN-PPARα Pathway

doi: 10.3390/nu18050872

Figure Lengend Snippet: The effect of PA on lipid accumulation in ALD mice. ( A ) Schematic of the acute alcoholic liver injury model in mice with propionate treatment. ( B , C ) Serum ALT and AST levels in mice. ( D , E ) H&E and Oil Red O staining of liver tissues (scale bar: 50 μm). ( F ) The relative area of Oil Red O staining. ( G , H ) Liver levels of TC and TG in mice. ( I – L ) Serum concentrations of TC, TG, HDL-C and LDL-C in mice. ( M ) Representative Western blot analysis and ( N ) quantification of ACC1, SREBP1 and CPT1A protein expression in liver tissues normalized to β-actin (n = 4). The data are presented as means ± SEM. * p < 0.05; ** p < 0.01.

Article Snippet: Primary antibodies, including β-actin (#66009-1-Ig), CPT1A (#15184-1-AP), SREBP-1 (#14088-1-AP), ACC1 (#21923-1-AP), ACOX1 (#83731-2-RR), PPARα (#66826-1-Ig) and RGN (#17947-1-AP), were obtained from Proteintech Group (Wuhan, China).

Techniques: Staining, Western Blot, Expressing

Journal: Nutrients

Article Title: Integrated Proteomics and Metabolomics Reveal the Direct Hepatic Protection of Propionate Against Alcoholic Liver Disease via the RGN-PPARα Pathway

doi: 10.3390/nu18050872

Figure Lengend Snippet: Propionate suppressed cell injury and lipid accumulation in vitro. ( A ) Changes in AML-12 cell viability after a series of doses of ethanol challenge. ( B ) Changes in AML-12 cell viability after a series of concentrations of OA incubation. ( C ) Effects of different concentrations of propionate on the viability of AML-12 cells. ( D ) Propionate restored AML-12 cells’ viability following co-treatment with 200 mM ethanol and 0.5 mmol/L OA. ( E ) Representative views of Oil Red O staining of AML-12 cells (scale bars: 100 μm and 50 μm). ( F ) Representative images of Western blot analysis and ( G ) quantification of ACC1, SREBP1 and CPT1A protein expression in AML-12 cells normalized to β-actin (n = 3). The data are presented as means ± SEM. * p < 0.05; ** p < 0.01.

Article Snippet: Primary antibodies, including β-actin (#66009-1-Ig), CPT1A (#15184-1-AP), SREBP-1 (#14088-1-AP), ACC1 (#21923-1-AP), ACOX1 (#83731-2-RR), PPARα (#66826-1-Ig) and RGN (#17947-1-AP), were obtained from Proteintech Group (Wuhan, China).

Techniques: In Vitro, Incubation, Staining, Western Blot, Expressing

Journal: Journal of Functional Biomaterials

Article Title: Cytocompatibility Assessment of L-PBF-Manufactured Zinc–Silver–Copper Alloys for Customized Biodegradable Medical Implants

doi: 10.3390/jfb17030146

Figure Lengend Snippet: ( A ) Representative images of the direct cytocompatibility contact test with SAOS-2 cells are presented. The top row ( a ) shows the tested sample plates (black) at the top of the image, with cells growing in direct contact on the bottom of the six-well plate (five times magnification). ( b ) The cell density at a 12 mm distance from the test plates (ten times magnification). ( B , D ) Three different extract concentrations (undiluted, diluted 1:3 and diluted 1:10) from the three L-PBF-manufactured Zn alloys (ZnAgCu, ZnAgCuMn, and ZnAgCuTi), rolled Zn, Cu, and Ti controls were tested, and the proliferation of L929 ( B ) and SAOS-2 ( D ) cells was determined after 24 h. ( C , E ) Live/dead staining of L929 ( C ) and SAOS-2 ( E ) cells growing directly on top of the samples is shown. Red and yellow staining indicate dead or dying cells, respectively, while living cells appear green. Each test was carried out four times with four replicates per sample. Shown are the mean values with their corresponding standard deviations. All results were normalized to control cells grown in cell culture medium without extract (proliferation equals 100%). To calculate statistical significance, a one-way ANOVA followed by a Tukey test was carried out (** p < 0.01 and **** p < 0.0001).

Article Snippet: In addition, the human primary osteogenic sarcoma cell line SAOS-2 (Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH), cultured in McCoy’s 5A medium (Sigma-Aldrich Chemie GmbH, Steinheim, Germany) supplemented with 15% FBS (Bio&SELL GmbH) and 1% L-glutamine (Thermo Fisher Scientific Inc.), was used.

Techniques: Staining, Control, Cell Culture

Journal: Journal of Functional Biomaterials

Article Title: Cytocompatibility Assessment of L-PBF-Manufactured Zinc–Silver–Copper Alloys for Customized Biodegradable Medical Implants

doi: 10.3390/jfb17030146

Figure Lengend Snippet: Time dependency of ion release and cytocompatibility of undiluted extracts tested for 24 h with SAOS-2 cells from untreated ( A ) and polished ( B ) L-PBF-manufactured plates. Extracts collected daily over a period of 10 days. ( C ) OM of untreated and polished plates before and after 10 days of immersion. The mean proliferation values with the corresponding standard deviation of independent experiments ( n = 4) are plotted graphically. All results were normalized to the control cells grown in cell culture medium without extract (=100%). Significance was determined using one-way ANOVA followed by a Tukey test (* p < 0.05; ** p < 0.01; *** p < 0.001; and **** p < 0.0001). ( D , E ) ICP-OES ion concentration measurement. ( D ) Zn 2+ release of the different alloys into daily renewed McCoy’s medium from day 1 to day 10. The extracts were taken from untreated and freshly polished samples. The mean values with the corresponding standard deviations of five independent experiments are plotted. ( E ) Ion concentrations of Zn 2+ , Ag + , Cu 2+ , and Mn 2+ release of ZnAgCuMn into the daily changed McCoy’s medium ( n = 5).

Article Snippet: In addition, the human primary osteogenic sarcoma cell line SAOS-2 (Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH), cultured in McCoy’s 5A medium (Sigma-Aldrich Chemie GmbH, Steinheim, Germany) supplemented with 15% FBS (Bio&SELL GmbH) and 1% L-glutamine (Thermo Fisher Scientific Inc.), was used.

Techniques: Standard Deviation, Control, Cell Culture, Concentration Assay

Journal: Journal of Functional Biomaterials

Article Title: Cytocompatibility Assessment of L-PBF-Manufactured Zinc–Silver–Copper Alloys for Customized Biodegradable Medical Implants

doi: 10.3390/jfb17030146

Figure Lengend Snippet: Cytocompatibility extract test with SAOS-2 cells cultivated for 24 h in extracts from untreated, new polished and aged polished samples. The mean proliferation values with the corresponding standard deviation of independent experiments ( n = 8) are plotted graphically. All results were normalized to the control cells grown in cell culture medium without extract (=100%). Significance was determined using one-way ANOVA followed by a Tukey test (* p < 0.05; ** p < 0.01).

Article Snippet: In addition, the human primary osteogenic sarcoma cell line SAOS-2 (Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH), cultured in McCoy’s 5A medium (Sigma-Aldrich Chemie GmbH, Steinheim, Germany) supplemented with 15% FBS (Bio&SELL GmbH) and 1% L-glutamine (Thermo Fisher Scientific Inc.), was used.

Techniques: Standard Deviation, Control, Cell Culture

Journal: Leukemia

Article Title: BET inhibitor-based combinations targeting novel dependencies in MECOM-rearranged (r) AML

doi: 10.1038/s41375-025-02842-w

Figure Lengend Snippet: A AML194 cells were treated with 500 nM of mivebresib as biologic replicates for 16 h. Total RNA was isolated and utilized for RNA-Seq analysis. Gene set enrichment analysis of mivebresib-treated over DMSO-control AML194 mRNA expression changes compared to HALLMARK pathways. Normalized enrichment scores are shown. All q-values are < 0.1. B UCSD-AML1 and AML194 cells were treated with 500 nM of mivebresib as biologic replicates for 16 h. Total RNA was isolated and utilized for cDNA generation and qPCR analysis. Messenger RNA expression is normalized to GAPDH and expressed relative to the DMSO control cells. Columns, mean of three replicates; Bars, standard error of the mean (S.E.M.). C AML194 cells were treated with 500 nM of mivebresib as biologic replicates for 24 h. Total proteome profiling was conducted by mass spectrometry analysis. Gene set enrichment analysis of mivebresib-treated over DMSO-control AML194 protein expression changes compared to HALLMARK, REACTOME and KEGG pathways is shown. Normalized enrichment scores are shown at a p -value < 0.01. D The volcano plot (log2 fold-change versus –log10 p -value) shows mass spectrometry-determined protein expression changes in mivebresib-treated compared to DMSO-treated control AML194 cells. E AML191, AML194, and UCSD-AML1 cells were treated with the indicated concentrations of mivebresib for 24 h. At the end of treatment, cells were harvested, total cell lysates were prepared, and immunoblot analyses were conducted. The expression levels of β-Actin in the total cell lysates served as the loading control. Representative blots of at least two experiments are shown.

Article Snippet: UCSD-AML1 [DSMZ Cat# ACC-691, RRID:CVCL_1853] and MUTZ-3 [DSMZ Cat# ACC-295 (RRID:CVCL_1433)] cells were obtained from the DSMZ (Braunschweig, Germany).

Techniques: Isolation, RNA Sequencing, Control, Expressing, RNA Expression, Mass Spectrometry, Western Blot

Journal: Leukemia

Article Title: BET inhibitor-based combinations targeting novel dependencies in MECOM-rearranged (r) AML

doi: 10.1038/s41375-025-02842-w

Figure Lengend Snippet: A UCSD-AML1, AML191 and AML194 cells were treated with mivebresib for 24 h. Following this, cells were fixed with 70% ethanol. Cell cycle status was determined by PI staining and flow cytometry. Columns, mean of three experiments; Bars, standard error of the mean (S.E.M.). * = p < 0.05 compared to DMSO-control cells as determined by a two-tailed, unpaired t-test in GraphPad V10 software. B AML191 and AML194 cells were treated with the indicated concentration of mivebresib for 96 h. Then, cells were stained with CD11b and CD86 antibodies and the % double-positive cells were analyzed by flow cytometry. C MUTZ-3, UCSD-AML1, AML191 and AML194 cells were treated with the indicated concentrations of mivebresib for 48 h. Following this, cells were stained with annexin V-FITC and TO-PRO-3 iodide and the % apoptotic cells were analyzed by flow cytometry. Lines indicate the mean of three experiments; Bars, standard error of the mean (S.E.M.). D, E Primary, patient-derived MECOM-r and non-MECOM-r AML samples and normal CD34+ HSPCs were treated with the indicated concentrations of mivebresib for 48 h. At the end of treatment, cells were stained with TO-PRO-3 iodide and analyzed by flow cytometry. * = p < 0.05 and ** = p < 0.01 compared to the non-MECOM-r AML cells as determined by a two-tailed, unpaired t-test in GraphPad V10 software.

Article Snippet: UCSD-AML1 [DSMZ Cat# ACC-691, RRID:CVCL_1853] and MUTZ-3 [DSMZ Cat# ACC-295 (RRID:CVCL_1433)] cells were obtained from the DSMZ (Braunschweig, Germany).

Techniques: Staining, Flow Cytometry, Control, Two Tailed Test, Software, Concentration Assay, Derivative Assay

Journal: Leukemia

Article Title: BET inhibitor-based combinations targeting novel dependencies in MECOM-rearranged (r) AML

doi: 10.1038/s41375-025-02842-w

Figure Lengend Snippet: A Table of target level, MECOM-r AML specific sensitivities identified from the drug screen shown by drug treatment set enrichment analysis (DTSEA)-determined normalized enrichment score with corresponding f.d.r. q -value < 0.1. B MUTZ-3, UCSD-AML1, AML191 and AML194 cells were treated with the indicated concentrations of dactolisib for 48 h. Following this, cells were stained with annexin V-FITC and TO-PRO-3 iodide and the % apoptotic cells were analyzed by flow cytometry. Lines indicate the mean of three experiments; Bars, standard error of the mean (S.E.M.). C , D Primary, patient-derived MECOM-r and non-MECOM-r AML or normal CD34 + HSPC samples were treated with the indicated concentrations of dactolisib for 48 h. At the end of treatment, cells were stained with TO-PRO-3 iodide and analyzed by flow cytometry. * = p < 0.05 compared to non-MECOM-r AML cells. E MUTZ-3, UCSD-AML1, AML191 and AML194 cells were treated with the indicated concentrations of LCL161 for 48 h. Then, cells were stained with annexin V-FITC and TO-PRO-3 and the % apoptotic cells were analyzed by flow cytometry. Lines indicate the mean of three experiments; Bars, standard error of the mean (S.E.M.). F , G Primary, patient-derived MECOM-r and non-MECOM-r AML or normal CD34 + HSPC samples were treated with the indicated concentrations of LCL161 for 48 h. Following this, the cells were stained with TO-PRO-3 iodide and analyzed by flow cytometry. * = p < 0.05; ** = p < 0.01. H Total bioluminescence flux (photons/second) in NSG mice engrafted with luciferase-expressing MECOM-r AML242 xenografts and treated with mivebresib (0.5 mg/kg), dactolisib (20 mg/kg) or LCL161 (65 mg/kg) for 3 weeks. ** = p < 0.01; *** = p < 0.005 as determined by a two-tailed, unpaired t-test in GraphPad V10 software. I. Kaplan-Meier survival plot of NSG mice engrafted with luciferase-expressing MECOM-r AML242 xenografts and treated for 4 weeks as indicated. * = p < 0.05; **** = p < 0.001 as determined by a log rank sum test in GraphPad V10 software.

Article Snippet: UCSD-AML1 [DSMZ Cat# ACC-691, RRID:CVCL_1853] and MUTZ-3 [DSMZ Cat# ACC-295 (RRID:CVCL_1433)] cells were obtained from the DSMZ (Braunschweig, Germany).

Techniques: Staining, Flow Cytometry, Derivative Assay, Luciferase, Expressing, Two Tailed Test, Software

Journal: Leukemia

Article Title: BET inhibitor-based combinations targeting novel dependencies in MECOM-rearranged (r) AML

doi: 10.1038/s41375-025-02842-w

Figure Lengend Snippet: A AML191, UCSD-AML1 and MUTZ-3 or AML194 cells were treated with mivebresib and/or dactolisib, LCL161, birinapant, bimiralisib, A1155463 or navitoclax for 48 h. At the end of treatment, cells were stained with annexin V-FITC and TO-PRO-3 iodide and the % apoptotic cells were analyzed by flow cytometry. Delta synergy scores were determined by the SynergyFinder web application using the ZIP method. Delta synergy scores greater than 1 indicate a synergistic interaction of the two drugs. Mean delta synergy scores are shown for each combination in each cell line. B , C AML194 cells were treated with the indicated concentrations of dactolisib or LCL161 and/or mivebresib for 24 h. Then, cells were harvested, total cell lysates were prepared and immunoblot analyses were conducted. The expression levels of β-Actin in the total cell lysates served as the loading control. Representative blots of at least two experiments are shown. Vertical lines indicate a repositioned gel image. Numbers beneath the blots indicate densitometry analysis compared to the DMSO-control cells.

Article Snippet: UCSD-AML1 [DSMZ Cat# ACC-691, RRID:CVCL_1853] and MUTZ-3 [DSMZ Cat# ACC-295 (RRID:CVCL_1433)] cells were obtained from the DSMZ (Braunschweig, Germany).

Techniques: Staining, Flow Cytometry, Western Blot, Expressing, Control