human bcl 2 polyclonal antibody  (Proteintech)

Journal: PLOS One

Article Title: Synergistic effects of notoginsenoside R1 and saikosaponin B2 in atherosclerosis: A novel approach targeting PI3K/AKT/mTOR pathway and macrophage autophagy

doi: 10.1371/journal.pone.0326687

Figure Lengend Snippet: (A) ox-LDL induces ROS generation in RAW264.7 cells, visualized by inverted fluorescence microscopy (scale bar = 50 μm). (B) Representative images of apoptosis detected by flow cytometry. (C) Representative Western blot images showing protein expression of BAX, BCL2, and Caspase-3 in the ox-LDL-induced cell model. β-actin was used as a loading control. (D-F) Quantification of the relative protein levels of BAX, BCL2, and Caspase-3 normalized to β-actin, and presented as relative intensity measured by Western blot analysis. All data were presented as the mean ± SD, (n = 3).** p < 0.01 vs. control group, ## p < 0.01 vs. model group, $$ p < 0.01 vs. NGR1 group, + p < 0.05, ++ p < 0.01 vs. SSB2 group.

Article Snippet: The antibodies included: AKT Monoclonal antibody (60203–2-Ig, Proteintech, China), Phospho-AKT (Ser473) Monoclonal antibody (66444–1-Ig, Proteintech, China), mTOR Monoclonal antibody (66888–1-Ig), Phospho-mTOR (Ser2448) Monoclonal antibody (67778–1-Ig, Proteintech, China), PI3 Kinase p110 Beta Polyclonal antibody (20584–1-AP, Proteintech, China), Phospho-PI3 Kinase p85 (Tyr458)/p55 (Tyr199) Antibody (4228T, CST, USA), IL-1 Beta Polyclonal antibody (16806–1-AP, Proteintech, China), IL-18 Polyclonal antibody (10663–1-AP, Proteintech, China), Nlrp3 Polyclonal antibody (30109–1-AP, Proteintech, China), Caspase 3/p17/p19 Polyclonal antibody (19677–1-AP, Proteintech, China), BAX Polyclonal antibody (50599–2-Ig, Proteintech, China), Bcl2 Polyclonal antibody (A00040-1, Boster, China), Anti-LC3B/MAP1LC3B Antibody (BM4827, Boster, China), Beclin-1 Antibody (3738T, CST, USA), Anti-P62/SQSTM1 Antibody (M00300-1, Boster, China).After incubation with secondary antibodies (BA1056, Boster, China) for 1 hour, protein bands were visualized using an enhanced ECL chemiluminescence reagent (P0018AM, Beyotime, China) and an Odyssey infrared imaging scanner (LI-COR Biosciences).

Techniques: Fluorescence, Microscopy, Flow Cytometry, Western Blot, Expressing, Control

Journal: PLOS One

Article Title: Synergistic effects of notoginsenoside R1 and saikosaponin B2 in atherosclerosis: A novel approach targeting PI3K/AKT/mTOR pathway and macrophage autophagy

doi: 10.1371/journal.pone.0326687

Figure Lengend Snippet: Representative images of Oil Red O staining showing lipid droplets in RAW264.7 cells. Cells were examined under a light microscope (scale bar = 100 μm). (B) Representative images of cell apoptosis detected by flow cytometry. (C-F) The use of gene-specific oligonucleotide primers for the real-time PCR analysis of PPAR-γ, LXR-α, ABCA1 , and ABCG1. (G) Representative Western blot images showing protein expression levels of P-mTOR, mTOR, NLRP3, BAX, BCL2, Caspase3, P62, BECLIN1, and LC3 in ox-LDL-induced cellular models. β-actin was used as the loading control. (H-O) Quantification of relative protein levels were normalized to β-actin and were presented as relative intensity. (P) Representative confocal microscopy images demonstrating the co-localization of p-mTOR (green) and Lamp-1 (red) in RAW264.7 macrophages (scale bar = 10 μm). All data were presented as the mean ± SD, (n = 3). ** p < 0.01 vs. control group, # p < 0.05, ## p < 0.01 vs. model group, $ p < 0.05, $$ p < 0.01 vs. NS group, + p < 0.05, ++ p < 0.01 vs. NS group. Rapamycin: 0.5 μM, 3-MA: 3 mM.

Article Snippet: The antibodies included: AKT Monoclonal antibody (60203–2-Ig, Proteintech, China), Phospho-AKT (Ser473) Monoclonal antibody (66444–1-Ig, Proteintech, China), mTOR Monoclonal antibody (66888–1-Ig), Phospho-mTOR (Ser2448) Monoclonal antibody (67778–1-Ig, Proteintech, China), PI3 Kinase p110 Beta Polyclonal antibody (20584–1-AP, Proteintech, China), Phospho-PI3 Kinase p85 (Tyr458)/p55 (Tyr199) Antibody (4228T, CST, USA), IL-1 Beta Polyclonal antibody (16806–1-AP, Proteintech, China), IL-18 Polyclonal antibody (10663–1-AP, Proteintech, China), Nlrp3 Polyclonal antibody (30109–1-AP, Proteintech, China), Caspase 3/p17/p19 Polyclonal antibody (19677–1-AP, Proteintech, China), BAX Polyclonal antibody (50599–2-Ig, Proteintech, China), Bcl2 Polyclonal antibody (A00040-1, Boster, China), Anti-LC3B/MAP1LC3B Antibody (BM4827, Boster, China), Beclin-1 Antibody (3738T, CST, USA), Anti-P62/SQSTM1 Antibody (M00300-1, Boster, China).After incubation with secondary antibodies (BA1056, Boster, China) for 1 hour, protein bands were visualized using an enhanced ECL chemiluminescence reagent (P0018AM, Beyotime, China) and an Odyssey infrared imaging scanner (LI-COR Biosciences).

Techniques: Staining, Light Microscopy, Flow Cytometry, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Control, Confocal Microscopy

Journal: Frontiers in Molecular Biosciences

Article Title: Shortwave Infrared-Emitting Theranostics for Breast Cancer Therapy Response Monitoring

doi: 10.3389/fmolb.2020.569415

Figure Lengend Snippet: Dox-ANCs induce cytotoxicity through activation of cell-death pathways. MDA-MB-231 cells (10 4 cells/well) were treated with an equivalent dose of 0.02 mg/mL Dox-ANCs or free Dox over 72 h. Viability was assessed via an MTT assay at times 0, 1, 2, 4, 8, 12, 24, 48, and 72 h of treatment, and the absorbance at 570 nm was normalized to untreated controls (A) . Data is expressed as mean ± SEM for n = 3. The cytotoxic effect of Dox-ANC was delayed in comparison to free Dox during the first 24 h. To assess the effect of Dox and Dox-ANCs on cellular pathways of cell death, an immunoblot was run from samples of MDA-MB-231 cells lysed after 12 and 24 h of treatment with Dox (Dox-ANCs or free Dox) or untreated. Qualitative analysis of the blots revealed elevated levels of cleaved PARP and decreased expression of Bcl2 in cells treated with Dox-ANC and free Dox compared to untreated control (B) . This indicates an active cell death process in response to the delivery of Dox.

Article Snippet: Antibodies used included rabbit anti-human Bcl-2 monoclonal antibody (Human Specific) (Clone D55G8; Cell Signaling, Danvers, MA) at 1:1000 dilution, rabbit anti-human cleaved PARP (Asp214) polyclonal antibody (Cell Signaling, Danvers, MA) at 1:1000 dilution, rabbit anti-human β-actin polyclonal antibody (Abcam, Cambridge, MA) at 1:3000, and goat anti-rabbit IgG HRP polyclonal antibody (Abcam, Cambridge, MA) at 1:5000 dilution.

Techniques: Activation Assay, MTT Assay, Comparison, Western Blot, Expressing, Control