l 6  (Bioss)

Journal: Science Advances

Article Title: A transient RNP compartment boosts translation and growth of mammalian oocytes

doi: 10.1126/sciadv.aej5359

Figure Lengend Snippet: ( A ) Oocyte at the cluster stage stained for nascent translation by Click chemistry with the puromycin analog OPP (OPP, right column), negative control condition previously treated with puromycin for 1 hour (+puromycin, bottom row). The Zb is visualized through the autofluorescence of its organelles at 488 nm (organelles, left column). ( B ) Top row: live cluster stage oocyte labeled with 6-FAM-dC-puromycin (green) to reveal nascent peptides as fluorescent foci (arrowhead) in the Zb. Bottom row: control oocyte not exposed to 6-FAM-dC-puromycin. MitoTracker (magenta) highlights the Zb. ( C ) Cluster stage oocyte labeled with anti–phospho-mTOR Ser 2448 (pmTOR; middle); the Zb is visualized through the autofluorescence of its organelles at 488 nm (organelles, left). The right panel shows the merge of organelles (pink) and pmTOR (blue) staining. ( D ) Quantification of nascent translation signal intensity at the cluster stage as in (A). n = 22 oocytes for OPP and n = 22 oocytes for OPP + puromycin; five independent experiments. ( E ) Quantification of Phospho-mTOR signal intensity at the cluster stage as in (C). n = 34 oocytes; four independent experiments. ( F ) Oocyte at the cluster stage labeled with anti–phospho-mTOR Ser 2448 , (right column) after 2 hours of treatment with nocodazole (Noco; bottom row) or its solvent DMSO (top row). The Zb is visualized by autofluorescence of its organelles at 488 nm (organelles, left column). ( G ) Quantification of fluorescence intensity at the cluster stage from (F). n = 39 oocytes for DMSO, 2 hours; n = 39 oocytes for nocodazole, 2 hours; five independent experiments. h, hours. ( H ) Oocyte diameter at the cluster, intermediary, and fully grown stages measured with quantitative phase imaging. n = 139 oocytes for cluster, n = 82 oocytes for intermediary, and n = 160 oocytes for fully grown; six independent experiments. ( I ) Oocyte dry mass at the cluster, intermediary, and fully grown stages measured with quantitative phase imaging. n = 139 oocytes for the cluster stage, n = 82 oocytes for the intermediary stage, and n = 160 oocytes for the fully grown stage; six independent experiments. ( J ) Oocyte protein density at the cluster, intermediary, and fully grown stages estimated from measurements in (H) and (I). n = 139 oocytes for cluster, n = 82 oocytes for intermediary, and n = 160 oocytes for fully grown; six independent experiments. ( K ) Simple linear regression of dry mass values from (I) plotted against diameter measurements from (H) for the cluster, intermediary, and fully grown stages. ( L ) Simple linear regression of density values from (J) plotted against diameter measurements from (H) for the cluster, intermediary, and fully grown stages. Data are shown as means ± SD. P values were calculated using unpaired two-tailed Student’s t test with Welch’s correction for (D), Mann-Whitney test for (G), or one-way ANOVA with Tukey’s post hoc test for (H), (I), and (J). Scale bars, 10 μm. n.s., not significant; * P < 0.05; ** P < 0.01; **** P < 0.0001.

Article Snippet: DND-26 LysoTracker Green (Thermo Fisher Scientific, L7526, stock solution in DMSO at 1 mM) was used at 50 nM for 20 min. 6-FAM-dC-puromycin (JenaBioscience, catalog number NU-925-6FM) was used at 50 μM for 10 min. Puromycin (Sigma-Aldrich, P7255) was diluted in water to make a stock solution at 50 mg/ml, used at 100 μg/ml for 1 hour.

Techniques: Staining, Negative Control, Labeling, Control, Solvent, Fluorescence, Imaging, Two Tailed Test, MANN-WHITNEY

Journal: Aging Cell

Article Title: Plasma Dilution After Myocardial Ischemia–Reperfusion Injury Promotes Cardiac Repair, Heart Performance, and Recovery of Motor Function and Endurance in Old Mice

doi: 10.1111/acel.70525

Figure Lengend Snippet: NBE restores de novo cardiac proteome levels to normal. (A) Schematic diagram. NBE was performed 24 h post‐injury. Mice were injected with azido‐homo‐alanine for 5 days starting with the onset of injury. Heart samples were harvested at 5 days post injury. (B) Principal component analysis (PCA) plots of the protein expression profile of all treatment groups showed clear separation of sham and I/R controls, with clustering of I/R + NBE group in sham. Confidence ellipses at 50%, 80%, and 95% intervals are overlaid to indicate group variance. (C) Volcano plots identified differentially expressed proteins (DEPs) in I/R versus Sham controls and I/R + NBE versus I/R controls. Proteins highlighted in red are significantly upregulated while proteins highlighted in blue are significantly downregulated (adjusted p ‐value < 0.05) (horizontal dotted line), and log2 fold change > 2 (vertical dotted line). (D) Venn diagram showed the 12 overlapping DEPs between the upregulated proteins after injury and their downregulation after NBE treatment. (E) Heatmap visualized the expression of the 12 overlapping proteins across all groups. (F, G) Gene Ontology overrepresentation analysis of the 12 overlapping DEPs highlighted top 10 biological processes and molecular function, respectively. (H) cirFunMap visualization of KEGG enriched pathways of the 12 overlapping proteins.

Article Snippet: After injury, mice were injected intraperitoneally with azido‐homo‐alanine (Jena Biosciences CLK‐AA009, 0.02 mmol/kg) daily for 5 days.

Techniques: Injection, Expressing