Journal: Developmental cell

Article Title: AAGAB controls AP2 adaptor assembly in clathrin-mediated endocytosis

doi: 10.1016/j.devcel.2019.06.013

Figure Lengend Snippet: KEY RESOURCE TABLE

Article Snippet: δ-adaptin , DSHB , Cat# anti-delta SA4, RRID:AB_2056641.

Techniques: Virus, Recombinant, Electron Microscopy, Protease Inhibitor, DNA Extraction, Mutagenesis, Sequencing, Library Amplification, CRISPR, Plasmid Preparation, Software

Journal: Frontiers in Cellular Neuroscience

Article Title: Role of a Pdlim5:PalmD complex in directing dendrite morphology

doi: 10.3389/fncel.2024.1315941

Figure Lengend Snippet: PalmD a novel partner of Pdlim5. (A) The image represents the colocalizations of Pdlim5 (shown in red), PalmD (shown in green) and DAPI (shown in blue) in rat hippocampal neurons at DIV4. Gross colocalization is indicated by the overlap color yellow in regions where both proteins are present within the same cellular compartment. Quantitation in Panel (B) . (B) The mean Pearson’s coefficient was calculated to quantify the extent of colocalization between Pdlim5 and PalmD in neurites. The value 0.65 suggests significant (but not complete) linear correlation of Pdlim5 and PalmD colocalization within neurites. Three biological replicates were evaluated with similar outcomes (one shown), with data from ≥ 15 neurons each. Error bars indicate the standard error of the mean (SEM). Each dot in the graph represents one neuron. (C) Immunofluorescent image displays the results of a proximity ligation assay (PLA) of rat hippocampal neurons at DIV4, where the red-colored puncta indicate the positive reaction produced when antibodies to Pdlim5 and PalmD were employed. White arrows point to examples of positive (red) puncta while F-actin staining images shown side by side for Pdlim5 and PalmD (negative controls, green). Quantitation in Panel (D) . (D) The quantification of PLA puncta per cell observed in processes is plotted. Antibodies employed against endogenous Pdlim5 or PalmD alone served as negative controls, while the established endogenous complex of Pdlim5:delta-Catenin served as positive control. The data was collected from ≥ 15 neurons, with each dot representing one neuron. Three biological replicates were evaluated with similar outcomes (one shown). The error bars represent the standard error of the mean (SEM), and the statistical significance, determined using one-way ANOVA, is indicated as P ≤ 0.0001 (****). ns, non significance. (E) The bar graphs compare the distribution of the Pdlim5 and PalmD proteins in different subcellular compartments, namely soma, neurites, and nucleus, in rat primary hippocampal neurons at two different developmental stages (DIV1 and DIV5). The relative fluorescence intensity for Pdlim5 and PalmD was quantified using ImageJ software. The data was obtained from 15 neurons, with each dot in the bar graph representing data from a single neuron. The error bars indicate the standard error of the mean (SEM). To determine the statistical significance, a one-way ANOVA analysis was performed using GraphPad Prism software. The levels of significance are indicated as **** for P ≤ 0.0001, n ≥ 15 neurons. Relative to the nucleus, this signifies high statistical significance of the observed subcellular localizations of both Pdlim5 and PalmD to neurites and soma in rat hippocampal neurons.

Article Snippet: Anti-delta-catenin mouse monoclonal , BD transduction laboratories , 611537.

Techniques: Quantitation Assay, Proximity Ligation Assay, Produced, Staining, Positive Control, Fluorescence, Software

Journal: Frontiers in Cellular Neuroscience

Article Title: Role of a Pdlim5:PalmD complex in directing dendrite morphology

doi: 10.3389/fncel.2024.1315941

Figure Lengend Snippet: Star methods: key resources table.

Article Snippet: Anti-delta-catenin mouse monoclonal , BD transduction laboratories , 611537.

Techniques: Cell Culture, Protease Inhibitor, Plasmid Preparation, Western Blot, Stripping, Microscopy, Transduction, Software

Journal: Bioengineered

Article Title: Knockdown of growth factor receptor bound protein 7 suppresses angiogenesis by inhibiting the secretion of vascular endothelial growth factor A in ovarian cancer cells

doi: 10.1080/21655979.2021.2005225

Figure Lengend Snippet: Knockdown of GRB7 in SKOV-3 cells decreases the expression of proteins associated with angiogenesis. HUVECs were seeded in the lower chamber of a transwell insert, and SKOV-3 cells transfected with negative control siRNA (NC) or siRNAs targeting GRB7 (siGRB7-1 or siGRB7-2) were seeded in the upper chamber of the transwell insert. These conditions are designated as HUVECs-NC, HUVECs-siGRB7-1, and HUVECs-siGRB7-2. After co-culture for 24 h, HUVECs were harvested for western blotting to measure the expression of VEGFR2, p-MEK1, p-ERK1/2, NOTCH1, and DLL4

Article Snippet: All primary antibodies were purchased from Proteintech Group, Inc., and their catalog numbers and dilutions were as follows: anti-GRB7 (Catalog number 10,045-1-Ig; 1:1000), anti-vascular endothelial growth factor A (VEGFA) (19,003-1-AP, 1:2000), anti-vascular endothelial growth factor receptor-2 (VEGFR2) (26,415-1-AP, 1:800), anti-mitogen-activated protein kinase kinase 1 (MAP2K1/MEK1) (28,930-1-AP, 1:3000), anti-extracellular signal-regulated kinases 1 and 2 (ERK1/2) (28,733-1-AP, 1:4000), anti-notch receptor 1 (NOTCH1) (20,687-1-AP, 1:1000), anti-delta-like canonical Notch ligand 4 (DLL4) (21,584-1-AP, 1:600), and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (10,494-1-AP, 1:6000).

Techniques: Expressing, Transfection, Negative Control, Co-Culture Assay, Western Blot

Journal: The Journal of Clinical Investigation

Article Title: Chloride homeostasis dysfunction drives hyperactivation of corticotropin-releasing factor-expressing neurons in the amygdala in stress-induced hypertension

doi: 10.1172/JCI195536

Figure Lengend Snippet: ( A and B ) Changes in MAP ( A ) and HR ( B ) measured by radiotelemetry before, during and after CUMS in WKY rats, BHRs and age-matched unstressed WKY rats and BHRs ( n = 6 rats in each group). Data are expressed as means ± SEM. * P < 0.05 for CUMS BHR versus unstressed BHRs, # P < 0.05 for CUMS BHR versus CUMS WKY, ^ P < 0.05 for CUMS WKY versus unstressed WKY. Repeated measures of 2-way ANOVA with Tukey’s post hoc test. ( C ) Representative immunofluorescence images showing CRF immunopositive neurons (green), delta-FosB immunopositive neurons (red), and CRF and delta-FosB double-positive neurons in the CeA of WKY, CUMS WKY, BHR, and CUMS BHR group of rats ( n = 5 rats in each group). ( D and E ) Summary data showing the numbers of CRF-positive neurons ( D ) and delta-FosB positive neurons ( E ) in the CeA of these 4 groups. ( F ) The percentage of CRF and delta-FosB double-positive neurons in the CeA of these 4 groups ( n = 5 rats in each group). Data are expressed as means ± SEM. ** P < 0.01, *** P < 0.001. One-way ANOVA with Tukey’s multiple comparison tests.

Article Snippet: Subsequently, the sections were incubated with a mouse anti-delta-FosB antibody (1:50, #sc-398595), a rabbit anti-CRF antibody (1:100, #A1122, Abclonal) or a rabbit anti-NKCC1 antibody (1:100, #13884, Proteintech) overnight at 4°C.

Techniques: Immunofluorescence, Comparison