beta site app cleaving enzyme 1 bace1  (Proteintech)

Journal: Journal of Neuroinflammation

Article Title: Far infrared light irradiation enhances Aβ clearance via increased exocytotic microglial ATP and ameliorates cognitive deficit in Alzheimer’s disease-like mice

doi: 10.1186/s12974-022-02521-y

Figure Lengend Snippet: FIR light did not influence the Aβ production but enhanced microglial Aβ phagocytosis. A The Western blot analysis of key molecules involved in the process of Aβ production. Expression level of B APPfl, C sAPPα, D Nicastrin, E BACE1 and F PS1 ( n = 5 per group). G Representative images of Aβ (6E10, red), microglia (Iba1, green) and nuclei (DAPI, blue) co-staining from APP/PS1 treated with or without FIR light. H Quantification of microglial cells within 20 μm from the Aβ plaque boundary ( n = 102 to 103 plaques per group) in the cerebral cortex. I Representative images of Aβ (6E10, white), microglia (Iba1, red), phagosome (CD68, green) and nuclei (DAPI, blue) co-staining in the cerebral cortex of APP/PS1 mice treated with or without FIR light. J Quantification of the percentage of phagosome area in Iba1 positive area ( n = 49–52 per group). K Quantification of the percentage of 6E10 + /CD68 + /Iba1 + co-staining area normalized to the total 6E10 + area ( n = 49–52 per group). L Representative images of the uptake of FAM-Aβ 1-42 . M Image quantification of the corresponding fluorescent intensity of FAM-Aβ 1-42 engulfed by microglia ( n = 6 per group). Data were means ± SEM, *** p < 0.001. N.S., not significant. CytoD: cytochalasin D

Article Snippet: The membrane was blocked by 5% nonfat milk at room temperature for 60 min and then incubated at 4 °C overnight with the following primary antibodies: mouse monoclonal anti-postsynaptic density protein-95 (PSD-95) antibody (1:1000, 75-028, NeuroMab), mouse monoclonal anti-synaptophysin (1:1000, MABN1193, EMD Millipore), rabbit polyclonal anti-APP (1:1000, AF6219, Beyotime), mouse monoclonal anti-Aβ (6E10) antibody that also recognized soluble APPα (sAPPα) (1:10,000, Sig-39300, Biolegend), rabbit polyclonal anti-nicastrin (1:1000, 14071-1-AP, Proteintech), rabbit polyclonal anti-beta-site-APP cleaving enzyme 1 (BACE1) (1:1000, AF6273, Beyotime), rabbit polyclonal anti-PS1 (1:1000, 16163-1-AP, Proteintech), or rabbit monoclonal anti-β-actin (1:1000, 8457 s, Cell Signaling Technology).

Techniques: Western Blot, Expressing, Staining

Journal: Frontiers in Pharmacology

Article Title: Berberine Reduces Aβ 42 Deposition and Tau Hyperphosphorylation via Ameliorating Endoplasmic Reticulum Stress

doi: 10.3389/fphar.2021.640758

Figure Lengend Snippet: Berberine reduces Aβ 42 production and deposition in APP/PS1 mice. (A) Representative IHC staining images and quantification of Aβ 42 in the CA1 region of the hippocampal tissue of mice. Scale bar is 100 μm. (B) The contents of Aβ 42 in the supernatant of hippocampal tissue homogenate of APP/PS1 mice were detected by ELISA. (C) Western blot detection of APP and CTF99 expression in the hippocampal tissues of the mice, and quantitative analysis. (D–G) Western blot detection of p-PERK, PERK, p-eIF2α, eIF2α, and BACE1 in the hippocampal tissues of mice, and quantitative analysis of p-PERK, p-eIF2α, and BACE1. Data were presented as means ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. WT; # p < 0.05, ## p < 0.001 vs. APP/PS1 without BBR.

Article Snippet: The primary antibodies used were as follows: β-actin (AB clonal, AC028, 1:3000), tau/ps404 (Abcam, ab30666, 1:1,000), tau/ps202 (Abcam, ab108387, 1:1,000), binding-immunoglobulin protein (Bip) (AB clonal, A0241, 1:1,000), p-GSK3β Y216 (AB clonal, AP0261, 1:1,000), GSK3β (CST, #12456,1:1,000); eIF2α (Abcam, ab169528, 1:1,000), p‐eIF2α Ser51 (CST, #3398, 1:1,000), PRKR-like endoplasmic reticulum kinase (PERK) (Abcam, ab65142, 1:1,000), p-PERK T980 (CST,#3179, 1:1,000), APP (AB clonal, A16265, 1:1,000), APP-C-terminal fragment 99 (CTF99) (AB clonal, A11019, 1:1,000), and beta-site APP cleaving enzyme-1 (BACE1) (AB clonal, A5266, 1:1,000).

Techniques: Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing

Journal: Frontiers in Pharmacology

Article Title: Berberine Reduces Aβ 42 Deposition and Tau Hyperphosphorylation via Ameliorating Endoplasmic Reticulum Stress

doi: 10.3389/fphar.2021.640758

Figure Lengend Snippet: Berberine inhibits tau hyperphosphorylation, as well as BACE1 production and eIF2α activation. (A) Western blot detection of p-eIF2α, eIF2α, and BACE1 in TG-induced HT22/APP cells with or without PBA or BBR and quantitative analysis. (B) Western blot detection of p-GSK3β Y216, GSK3β, tau ps 202, and tau ps 404 in TG-induced HT22/APP cells with or without PBA or BBR and quantitative analysis. Data were presented as means ± SEM. * p < 0.05, ** p < 0.01 vs. Blank; # p < 0.05 vs. TG.

Article Snippet: The primary antibodies used were as follows: β-actin (AB clonal, AC028, 1:3000), tau/ps404 (Abcam, ab30666, 1:1,000), tau/ps202 (Abcam, ab108387, 1:1,000), binding-immunoglobulin protein (Bip) (AB clonal, A0241, 1:1,000), p-GSK3β Y216 (AB clonal, AP0261, 1:1,000), GSK3β (CST, #12456,1:1,000); eIF2α (Abcam, ab169528, 1:1,000), p‐eIF2α Ser51 (CST, #3398, 1:1,000), PRKR-like endoplasmic reticulum kinase (PERK) (Abcam, ab65142, 1:1,000), p-PERK T980 (CST,#3179, 1:1,000), APP (AB clonal, A16265, 1:1,000), APP-C-terminal fragment 99 (CTF99) (AB clonal, A11019, 1:1,000), and beta-site APP cleaving enzyme-1 (BACE1) (AB clonal, A5266, 1:1,000).

Techniques: Activation Assay, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Berberine Reduces Aβ 42 Deposition and Tau Hyperphosphorylation via Ameliorating Endoplasmic Reticulum Stress

doi: 10.3389/fphar.2021.640758

Figure Lengend Snippet: Berberine decreases Aβ 42 production by inhibiting eIF2α activation, but eIF2α had little effect to GSK3β. (A) Western blot detection of eIF2α and p-eIF2α in HT22 cells with si-eIF2α (siRNA eIF2α) or scramble and quantitative analysis. (B) Western blot detection of BACE1, p-GSK3β Y216, and GSK3β in TG-induced HT22/APP cells with or without si-eIF2α (siRNA eIF2α) or BBR and quantitative analysis. (C) The contents of Aβ 42 in TG-induced HT22/APP cells with or without si-eIF2α (siRNA eIF2α) or BBR were detected by ELISA. (D) Western blot detection of tau ps 202 and tau ps 404 in TG-induced HT22/APP cells with or without si-eIF2α (siRNA eIF2α) or BBR and quantitative analysis. (E) Western blot detection of p-GSK3β Y216, GSK3β, tau ps 202, and tau ps 404 in TG-induced HT22/APP cells with or without SB216763 (GSK3β inhibitor) or BBR and quantitative analysis. Data were presented as means ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. Blank; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. TG, & p < 0.05 vs. BBR.

Article Snippet: The primary antibodies used were as follows: β-actin (AB clonal, AC028, 1:3000), tau/ps404 (Abcam, ab30666, 1:1,000), tau/ps202 (Abcam, ab108387, 1:1,000), binding-immunoglobulin protein (Bip) (AB clonal, A0241, 1:1,000), p-GSK3β Y216 (AB clonal, AP0261, 1:1,000), GSK3β (CST, #12456,1:1,000); eIF2α (Abcam, ab169528, 1:1,000), p‐eIF2α Ser51 (CST, #3398, 1:1,000), PRKR-like endoplasmic reticulum kinase (PERK) (Abcam, ab65142, 1:1,000), p-PERK T980 (CST,#3179, 1:1,000), APP (AB clonal, A16265, 1:1,000), APP-C-terminal fragment 99 (CTF99) (AB clonal, A11019, 1:1,000), and beta-site APP cleaving enzyme-1 (BACE1) (AB clonal, A5266, 1:1,000).

Techniques: Activation Assay, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: Berberine Reduces Aβ 42 Deposition and Tau Hyperphosphorylation via Ameliorating Endoplasmic Reticulum Stress

doi: 10.3389/fphar.2021.640758

Figure Lengend Snippet: A functional model of berberine-protecting effects against ER stress induced production of senile plaques and hyperphosphorylation of tau in AD. In the APP/PS1 AD model, ER stress was induced, and subsequently activated GSK3β cascade and the PERK/eIF2α/BACE1 signaling pathway. Tau hyperphosphorylation and Aβ 42 deposition were attenuated by BBR, which is mainly attributed to its ability to suppress ER stress in the AD model.

Article Snippet: The primary antibodies used were as follows: β-actin (AB clonal, AC028, 1:3000), tau/ps404 (Abcam, ab30666, 1:1,000), tau/ps202 (Abcam, ab108387, 1:1,000), binding-immunoglobulin protein (Bip) (AB clonal, A0241, 1:1,000), p-GSK3β Y216 (AB clonal, AP0261, 1:1,000), GSK3β (CST, #12456,1:1,000); eIF2α (Abcam, ab169528, 1:1,000), p‐eIF2α Ser51 (CST, #3398, 1:1,000), PRKR-like endoplasmic reticulum kinase (PERK) (Abcam, ab65142, 1:1,000), p-PERK T980 (CST,#3179, 1:1,000), APP (AB clonal, A16265, 1:1,000), APP-C-terminal fragment 99 (CTF99) (AB clonal, A11019, 1:1,000), and beta-site APP cleaving enzyme-1 (BACE1) (AB clonal, A5266, 1:1,000).

Techniques: Functional Assay

Journal: PLoS ONE

Article Title: Hook Proteins: Association with Alzheimer Pathology and Regulatory Role of Hook3 in Amyloid Beta Generation

doi: 10.1371/journal.pone.0119423

Figure Lengend Snippet: (a) N2A cells were transfected with two different siRNAs targeting Hook3 and with non-target siRNA as control (n = 3). Beta-amyloid accumulated for 48h in cell culture media was quantified by a murine amyloid-specific ELISA. Hook3 downregulation by two different siRNAs causes a 35% increase in secreted β-amyloid. * p < 0.05 compared to control group with student’s t-test. Data are means ± SD. (b) Western blot analysis of cell lysates demonstrate that transfection with Hook3 siRNA does not alter the levels of intracellular APP, BACE1 and PS1.

Article Snippet: Additionally, we used mouse antibodies against dynein intermediate chain (DIC): 74–1 (Santa Cruz), α-tubulin (DM1A), β-actin (AC-74) and S100β (SH-B1, all Sigma), p150 and p50 (BD Transduction Laboratories), pan-tau (clone 8F10, Analytik Jena), phospho-tau (AT8 and AT100, Thermo Scientific), APP (clone 22C11 Chemicon), β-amyloid (6F/3D, Dako) and rabbit antibodies against kinesin light chain 1 (KLC1, Santa Cruz), beta-site APP-cleaving enzyme 1 (BACE1, Calbiochem), ionized calcium binding adaptor molecule 1 (IBA1, Wako Chemicals), glial fibrillary acidic protein (GFAP, Dako), pan-tau (A0024, Dako) and goat anti-Presenilin 1 (PS1 N-19, Santa Cruz).

Techniques: Transfection, Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot