Journal: International Journal of Molecular Sciences

Article Title: Role of PI3 Kinases in Cell Signaling and Soleus Muscle Atrophy During Three Days of Unloading

doi: 10.3390/ijms26010414

Figure Lengend Snippet: Evaluation of MuRF1 ( A ), MAFbx ( B ), ubiquitin ( C ), and TFEB ( D ) mRNA expression and phospho-FOXO3 ( E ) content in the muscles of control (3C), three-day unloaded (3HS), and LY294002-treated three-day unloaded (3LY) rats. Phosphorylated FOXO3 content was normalized to the total FOXO3 content in each sample ( E ). N = 8. * indicates a significant difference from the control, and # indicates a significant difference from the 3HS group, p < 0.05.

Article Snippet: The following primary antibodies were used: IP3 receptors (1:500, #PA5-96855), phospho-CaMKII (Thr286) (1:500, #PA1-4614), phospho-FoxO3 (Ser253) (1:1000, #PA5-37578), and FoxO3 (1:1000, #PA5-20973) from Thermo Fisher Scientific, Waltham, MA, USA.

Techniques: Ubiquitin Proteomics, Expressing, Muscles, Control

Journal: Nature Communications

Article Title: Membrane compression by synaptic vesicle exocytosis triggers ultrafast endocytosis

doi: 10.1038/s41467-023-38595-2

Figure Lengend Snippet: a (top) Example airyscan micrographs showing the localization of Epsin1 relative to F-actin in wild-type neurons. Epsin1 stained with Epsin1-antibody and its secondary antibody conjugated with Atto647N. F-actin is labeled by neuronal expression of EGFP-UtrCH, which is stained by a GFP-antibody and its secondary antibody conjugated to Atto488. (bottom) Plot showing the Pearson’s correlation coefficient. Each dot = synapse. n = 94 synapses. N = 3 cultures. b Example 2D STED micrographs showing the localization of Epsin 1 relative to the active zone in wild-type neurons. Active zone is marked by anti-Bassoon antibody and its secondary antibody conjugated with Alexa594. Epsin1 stained with Epsin1-antibody and its secondary antibody conjugated with Atto647N. c The distribution of Epsin1 signals against the active zone boundary. The active zone boundary was defined by Bassoon signals. See Methods for the analysis method. Error bars are SEM. N = 2 cultures, n = 30–60 per culture. d Example confocal fluorescence micrographs showing F-actin signals in neurons expressing scramble shRNA and Epsin1 knock-down (KD) shRNA. F-actin is stained by expressing EGFP-UtrCH. e The normalized intensity of F-actin signals from neurons expressing scramble (scr) shRNA, or Epsin1 shRNA, measured by Airyscan. Signals are normalized to the fluorescence signals in axons. Kruskal-Wallis test, with Dunn’s multiple comparisons test. *** p < 0.001. See Supplementary Table for the detailed numbers for each sample. f Example electron micrographs showing wild-type and Epsin1 KD synapses unstimulated or stimulated with a single electrical pulse (1 ms) and frozen 100 ms or 1 s later. Black arrow: endocytic pit. Black arrowhead: ferritin-positive endosomes. g , h Number of endocytic pits at 100 ms after stimulation ( g ) or ferritin-positive structures at 1 s after stimulation ( h ) in neurons expressing scramble shRNA (scrRNA) or Epsin1 shRNA. Mean and 95% confidential interval are shown. Kruskal-Wallis test, with Dunn’s multiple comparisons test. **** p < 0.0001. p-values are only shown for direct comparison between unstimulated and stimulated neurons treated with the same drug. See Supplementary Table for the detailed numbers for each sample. Source data are provided as a Source Data file.

Article Snippet: For visualizing Epsin1, we used a plasmid expressing Epsin1-EGFP, obtained from Addgene (pEGFPC1-Epsin1, #22228).

Techniques: Staining, Labeling, Expressing, Fluorescence, shRNA, Knockdown, Comparison

Journal: Cell Reports

Article Title: The transcription factor ZEB1 regulates stem cell self-renewal and cell fate in the adult hippocampus

doi: 10.1016/j.celrep.2021.109588

Figure Lengend Snippet:

Article Snippet: Rabbit anti-INSC , Proteintech , Cat# 20973-1-AP; RRID: AB_10951111.

Techniques: Virus, Plasmid Preparation, Expressing, Recombinant, Software, Microscopy