Journal: Frontiers in Molecular Neuroscience

Article Title: Amyloid β1-42 (Aβ1-42) Induces the CDK2-Mediated Phosphorylation of Tau through the Activation of the mTORC1 Signaling Pathway While Promoting Neuronal Cell Death

doi: 10.3389/fnmol.2017.00229

Figure Lengend Snippet: Aβ facilitates HIF1α synthesis and autophagy inhibition via mTOR activation. (A) SK-N-MC cells were exposed to Aβ (5 μM) for 0–48 h. HIF1α and β-actin expression was analyzed by western blot. n = 3. (B) Cells were pretreated with NAC (1 mM) for 30 min prior to Aβ treatment for 24 h. HIF1α and β-actin expression were analyzed by western blot. n = 3. (C,E) Cells were incubated with rapamycin (10 nM) for 30 min prior to Aβ treatment for 24 h. Phosphorylation of 4EBP1 (Thr 37/46) and 4EBP1, phosphorylation of p70S6K1 (Thr 389), HIF1α and β-actin were analyzed by western blot. n = 6. (D) Protein samples were immunoprecipitated by eukaryotic translation initiation factor 4E (eIF4E) antibody-conjugated protein A/G agarose beads. Samples were blotted with 4EBP1 and eIF4E-specific antibodies. n = 3. (F) Cells were exposed to PF4708671 (10 μM) for 30 min prior to Aβ treatment for 24 h. HIF1α and β-actin expression was detected by western blot. n = 6. (G) Cells were exposed to cycloheximide (4 μM) for 30 min prior to Aβ treatment for 24 h. HIF1α and β-actin expressions were detected by western blot. n = 6. (H) Cells were pretreated with rapamycin (10 nM) for 30 min, incubated with Aβ for 24 h and analyzed by western blotting with LC3, p62 and β-actin specific antibodies. n = 3–6. (I) LC3 puncta was visualized by confocal microscopy. Presented results are merged images. Green and red fluorescents indicate LC3 and PI respectively. Scale bars, 50 μm (magnification × 600). (J) Cells were pretreated with trehalose (10 μM) for 30 min prior to Aβ treatment for 24 h. Cytotoxicity was measured by MTT assay at an absorbance of 545 nm using a microplate reader. Data present the mean ± SE. n = 6. (K) Cell viability was measured by trypan blue exclusion assay. Data are presented as a mean ± SE. n = 6. Each blot image was presented as representative image. * p < 0.05 vs. control, # p < 0.05 vs. Aβ treatment.

Article Snippet: The antibodies of hypoxia inducible factor (HIF1α; NB100-105), p70S6K1 (NB600-1049), LC3 (NB100-2220) and p62 (NBP1-48320) were obtained from Novus Biologicals (Littleton, CO, USA) and the HRP-conjugated goat anti-rabbit IgG was purchased from Santa Cruz Biotechnology.

Techniques: Inhibition, Activation Assay, Expressing, Western Blot, Incubation, Phospho-proteomics, Immunoprecipitation, Confocal Microscopy, MTT Assay, Trypan Blue Exclusion Assay, Control

Journal: International Journal of Biological Sciences

Article Title: Sesn2 Serves as a Regulator between Mitochondrial Unfolded Protein Response and Mitophagy in Intervertebral Disc Degeneration

doi: 10.7150/ijbs.70211

Figure Lengend Snippet: Antibody information

Article Snippet: Anti-LC3 , Novus , Cat. No. NB100-2220.

Techniques:

Journal: Heliyon

Article Title: Repurposing fluphenazine as an autophagy modulator for treating liver cancer

doi: 10.1016/j.heliyon.2023.e22605

Figure Lengend Snippet: Fluphenazine blocked autophagy flux of HCC cells. (A) The expression of autophagy related proteins was evaluated using Western blotting analysis in HepG-2 and MHCC97-H cells after treatment of fluphenazine for 48 h. Quantified values were shown on the right. (B) Confocal images of LC3B expression in HepG-2 and MHCC97-H cells treated with fluphenazine for 48 h. Nuclei were stained with DAPI. (C) Fluorescence images of LysoTracker Red in HepG-2 and MHCC97-H cells treated with fluphenazine for 48 h. Nuclei were stained with DAPI. (D) The cell viabilities of HepG-2 cells treated with 5 μM fluphenazine in the presence or absence of 1 mM 3-MA or 5 μM CQ for 48 h. (E) The cell viabilities of MHCC97-H cells treated with 10 μM fluphenazine in the presence or absence of 1 mM 3-MA or 5 μM CQ for 48 h. Data represent mean ± SD from at least three independent experiments (* P < 0.05, ** P < 0.01).

Article Snippet: Then the cells were incubated with LC3B antibody (Novus, NBP2-46892, 1:100) overnight at 4 °C, and washed with PBS for 5 min three times.

Techniques: Expressing, Western Blot, Staining, Fluorescence

Journal: Viruses

Article Title: Cyclin-Dependent Kinases 8 and 19 Regulate Host Cell Metabolism during Dengue Virus Serotype 2 Infection

doi: 10.3390/v12060654

Figure Lengend Snippet: Cell Protein Antibodies.

Article Snippet: Rabbit anti LC3B , Novus Biologicals , NBP246892SS.

Techniques: