Journal: Journal of Ovarian Research

Article Title: MSCs–derived EVs protect against chemotherapy-induced ovarian toxicity: role of PI3K/AKT/mTOR axis

doi: 10.1186/s13048-024-01545-7

Figure Lengend Snippet: Effect of MSCs-EVs, rapamycin, and quercetin on ( A ) phosphorylation of mTOR, PI3K, and AKT as detected by western blot analysis, ( B - D ) Intensity of immunoreactivity of selected proteins as quantified by densitometry. Results are expressed as mean ± SEM. **significant vs. Control group at p<0.01, **** at p<0.0001, # significant vs. OF group at p<0.05, ## at p<0.01, ### at p<0.001, #### at p<0.0001, & significant vs. OF + Rapamycin group at p<0.05

Article Snippet: Next, the blots were incubated with the appropriate primary antibodies for an entire night at 4 °C, PTEN (E-AB-63495, Elabscience, USA), FOXO3 (NBP2-16521, Novus Biologicals USA), mTOR (sc-517464, Santa Cruz Biotechnology, USA), Phospho-mTOR (sc-293133, Santa Cruz Biotechnology, USA), PI3K (E-AB-64202, Elabscience, USA), Phospho-PI3K (E-AB-20966, Elabscience, USA), AKT (E-AB-15441, Elabscience, USA), Phospho-AKT (E-AB-20804, Elabscience, USA), β-actin (E-AB-20031, Elabscience, USA).

Techniques: Phospho-proteomics, Western Blot, Control

Journal: Neurochemical Research

Article Title: Morin Improves Cognitive Deficits in an in Vivo Model of Vascular Dementia by Modulating the N-methyl-D-aspartate Receptor Signaling Pathways

doi: 10.1007/s11064-026-04717-7

Figure Lengend Snippet: Morin modulated the expression of NMDA receptors in the hippocampus of VaD rats. a the expression levels of NR1 ; b the expression levels of NR2A ; c the expression levels of NR2B ; d the expression levels of NR1 protein; e the expression levels of NR2A protein; f the expression levels of NR2B protein; g protein levels of p-CREB; h protein levels of p-CAMK2A; i protein levels of p-CAMK2D. Protein levels of NR1, NR2A, and NR2B were quantified by ELISA. Data are presented as mean ± SD ( n = 8 per group). Statistical analysis was performed by one-way ANOVA with Tukey’s post-hoc test (data met assumptions of normality and homoscedasticity)/Kruskal-Wallis with Dunn’s test (data did not meet assumptions). * indicates a significant difference from the Sham group; # indicates a significant difference 2VO group; *# indicates a significant difference from both the Sham and 2VO groups, with a p-value of less than 0.05 considered statistically significant

Article Snippet: Moreover, phosphorylation levels of calcium/calmodulin-dependent protein kinase II isoforms CAMK2A and CAMK2D at Thr286 (p-CAMK2A, p-CAMK2D) were quantified using the Colorimetric Cell-Based ELISA Kit (CAMK2A/CAMK2D (Phospho-Thr286), Boster Bio, #EKC2366).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Cell

Article Title: A Flat BAR Protein Promotes Actin Polymerization at the Base of Clathrin-Coated Pits

doi: 10.1016/j.cell.2018.05.020

Figure Lengend Snippet: FCHSD2 Is Not Directly Recruited to CCPs by PI3KC2α or Its Kinase Activity, Related to Figure 2 (A) Top: Transferrin uptake assay by flow cytometry comparing wild-type and FCHSD2 KO cells silenced for PI3KC2α. Uptake of Alexa488 labeled transferrin normalized by the amount of surface transferrin receptor for each condition and against uptake for the wild-type cells in each experiment. Each value represents median fluorescence from at least 5000 cells (n = 12, mean ± SD). ∗∗∗ p > 0.001, ns = non-significant. One-way ANOVA with Tukey’s post hoc analysis. Bottom: Immunoblots showing PI3KC2α knockdown in wild-type and FCHSD2 KO cells. (B) Kymographs of HeLa FCHSD2-Venus stables silenced for PI3KC2α and control cells. Cells were transfected with mCherry-ClathrinLC 24 hs before imaging. Kymographs generated from 120 s movies at 1Hz. Note the elongated CCP lifetimes in PI3KC2α knockdown cells as described in Posor et al. (2013) . (C) Autoinhibition of FCHSD2. Representative images of a center slice from cells expressing different FCHSD2 truncation constructs and co-stained with phalloidin (Actin). The bar graph (upper right) shows the quantification of cellular protrusions/μm for each construct. The non-inhibited BAR domain produces many protrusions. Numbers inside bars represent number of cells measured. The line graph (bottom right) shows the fluorescence profile of sum intensity projections for cells expressing each construct. Due to the natural thinning of cells from their centers to the edge, a gradually decaying line indicates that the fluorescent protein is primarily cytosolic while a flat line with an abrupt fall on the cell edge indicates that the fluorescent protein is primarily bound to the membrane. While the presence of SH3-1 significantly reduces the generation of cellular protrusions generated by the FCHSD2 F-BAR, a significant fraction of the protein remains bound to the membrane (green line). Only the combined presence of SH3-1 and SH3-2 is capable to avoid promiscuous binding of the BAR domain to the membrane. Data is shown as mean ± SD in bar graph and as ± SEM in fluorescence profiles. ∗∗∗ p > 0.001, ns = non-significant. One-way ANOVA with Tukey’s post hoc analysis. (D) Immunoblots for intersectin knockdown in FCHSD2-Venus HeLa cells.

Article Snippet: PI3KC2α siRNA pool , Origene , Cat# SR303516.

Techniques: Activity Assay, Flow Cytometry, Labeling, Fluorescence, Western Blot, Knockdown, Control, Transfection, Imaging, Generated, Expressing, Construct, Staining, Membrane, Binding Assay

Journal: Cell

Article Title: A Flat BAR Protein Promotes Actin Polymerization at the Base of Clathrin-Coated Pits

doi: 10.1016/j.cell.2018.05.020

Figure Lengend Snippet:

Article Snippet: PI3KC2α siRNA pool , Origene , Cat# SR303516.

Techniques: Virus, Recombinant, Labeling, Cloning, Stable Transfection, Expressing, esiRNA, Plasmid Preparation, Software