methotrexate (mts) solution  (Promega)

Journal: eLife

Article Title: JAK-STAT pathway activation compromises nephrocyte function in a Drosophila high-fat diet model of chronic kidney disease

doi: 10.7554/eLife.96987

Figure Lengend Snippet: ( A ) Representative confocal images of 7-day-old female adult nephrocytes ( 10xStat92E -GFP). Control, incubated in Schneider’s Drosophila Medium (ex vivo for 60 min at room temperature); methotrexate, incubated in 10 µM methotrexate in Schneider’s Drosophila Medium (ex vivo for 60 min at room temperature). 10xStat92E -GFP in green fluorescence. DAPI staining in blue to visualize the nucleus. Scale bar: 20 µm. ( B ) Box plot shows the quantitation of the relative fluorescence intensity of 10xStat92E -GFP based on the images in ( A ); middle line depicts the median and whiskers show Tukey. Statistical analysis was performed with a two-tailed t-test; ** p <0.01; n=6 flies.

Article Snippet: The dorsal cuticle (with nephrocytes) was transferred to methotrexate solution (10 μM) in Schneider’s Drosophila Medium (Thermo Fisher Scientific, MA) and incubated at room temperature for 60 min.

Techniques: Control, Incubation, Ex Vivo, Fluorescence, Staining, Quantitation Assay, Two Tailed Test

Journal: eLife

Article Title: JAK-STAT pathway activation compromises nephrocyte function in a Drosophila high-fat diet model of chronic kidney disease

doi: 10.7554/eLife.96987

Figure Lengend Snippet: Nephrocytes from control Drosophila ( w 1118 ; 7-day-old females) fed a regular diet (normal fat diet, NFD) or high-fat diet (NFD supplemented with 14% coconut oil, HFD), with or without methotrexate (10 μM; ex vivo 60 min) treatment. ( A ) Representative confocal images of FITC-albumin fluorescence (green). Scale bar: 50 μm. ( B ) Box plot shows the quantitation of the relative fluorescence intensity of FITC-albumin uptake based on images in ( A ); middle line depicts the median and whiskers show minimum to maximum. Statistical analysis was performed by two-way ANOVA with Sidak correction; *** p <0.001, **** p <0.0001; ns, not significant; n=6 flies (7-day-old females). ( C ) Representative confocal images of 10 kD dextran fluorescence (red). Scale bar: 50 μm. ( D ) Box plot shows the quantitation of the relative fluorescence intensity of 10 kD dextran uptake based on images in ( C ); middle line depicts the median and whiskers show minimum to maximum. Statistical analysis was performed by two-way ANOVA with Sidak correction; **** p <0.0001; ns, not significant; n=6 flies (7-day-old females). ( E ) Graphic of proposed model for high-fat diet-induced nephrocyte defects via an adipose-nephrocyte axis. A high-fat diet upregulates the expression and secretion of the adipokine Unpaired 2 (Upd2), leptin-like hormone, from the fat body. Upd2 is a Janus kinase/signal transducer and activator of transcription (JAK-STAT) ligand, and it activates JAK-STAT signaling at the nephrocytes (Signal-transducer and activator of transcription 92E, Stat92E; Suppressor of cytokine signaling at 36E, Socs36E; JAK Hopscotch, Hop; Domeless, Dome). The overactive JAK-STAT pathway disrupts the integrity of the slit diaphragm (SD) filtration structure and thereby leads to nephrocyte dysfunction.

Article Snippet: The dorsal cuticle (with nephrocytes) was transferred to methotrexate solution (10 μM) in Schneider’s Drosophila Medium (Thermo Fisher Scientific, MA) and incubated at room temperature for 60 min.

Techniques: Control, Ex Vivo, Fluorescence, Quantitation Assay, Expressing, Filtration

Journal: eLife

Article Title: JAK-STAT pathway activation compromises nephrocyte function in a Drosophila high-fat diet model of chronic kidney disease

doi: 10.7554/eLife.96987

Figure Lengend Snippet: ( A ) Representative confocal images of nephrocytes from control Drosophila ( sns-mRuby3 ; 7-day-old females) fed a regular diet (normal fat diet, NFD) or high-fat diet (NFD supplemented with 14% coconut oil, HFD), with or without methotrexate (10 μM; ex vivo 60 min) treatment. Sns-mRuby3 is in red. Scale bar: 5 μm. ( B ) Box plot shows the quantitation of Sns-mRuby3 protein distribution (cytoplasmic vs membrane) based on images in ( A ); middle line depicts the median and whiskers show minimum to maximum. Statistical analysis was performed by two-way ANOVA with Sidak correction; **** p <0.0001; ns, not significant; n=12 flies (7-day-old females).

Article Snippet: The dorsal cuticle (with nephrocytes) was transferred to methotrexate solution (10 μM) in Schneider’s Drosophila Medium (Thermo Fisher Scientific, MA) and incubated at room temperature for 60 min.

Techniques: Control, Ex Vivo, Quantitation Assay, Membrane

Journal: Scientific Reports

Article Title: Immunoprognostic model of lung adenocarcinoma and screening of sensitive drugs

doi: 10.1038/s41598-022-11052-8

Figure Lengend Snippet: CMap database drug screening.

Article Snippet: We further prepared a solution of resveratrol (Lot#:C12594500, Macklin, China), methotrexate (Lot#: C12150088, Macklin, China), phenoxybenzamine (Lot#: C11947938, Macklin, China), and thiostrepton.

Techniques:

Journal: Scientific Reports

Article Title: Immunoprognostic model of lung adenocarcinoma and screening of sensitive drugs

doi: 10.1038/s41598-022-11052-8

Figure Lengend Snippet: CMap database drug screening and cytological activity validation. ( a ) Optimal compounds and their structures screened from the CMap database. ( b ) DMSO solvent toxicity assay. ( c ) Inhibition rate of the resveratrol compound on A549 cells. ( d ) Inhibition rate of the methotrexate compound on A549 cells. ( e ) Inhibition rate of the phenoxybenzamine compound on A549 cells.

Article Snippet: We further prepared a solution of resveratrol (Lot#:C12594500, Macklin, China), methotrexate (Lot#: C12150088, Macklin, China), phenoxybenzamine (Lot#: C11947938, Macklin, China), and thiostrepton.

Techniques: Drug discovery, Activity Assay, Biomarker Discovery, Solvent, Inhibition