Journal: bioRxiv

Article Title: Development of a high-throughput, quantitative platform using human cerebral organoids to study virus-induced neuroinflammation in Alzheimer’s disease

doi: 10.1101/2024.03.21.585957

Figure Lengend Snippet: (A) UMAP plots showing the normalized fluorophore intensities for Aβ monomers using Solanezumab (red), HSV-1 (green) and Zombie fluorophore (blue) across uninfected, HSV-1 infected and ACV-treated cells. (B) Pairwise adjusted correlation heatmaps using the intensities for HSV-1, Zombie fluorophore and Solanezumab (Aβ monomers) across uninfected, HSV-1 infected and ACV-treated cells, shown for 2 sets of replicates (Replicate 1 and Replicate 2). (C) Boxen plots showing the normalized Aβ42 fluorophore intensities for uninfected cells (HSV-1 - cells) versus infected cells (HSV-1 + cells) within the same infected sample, as well as uninfected cells versus infected cells within the same ACV-treated sample. (D) Boxen plots showing the normalized Solanezumab fluorophore intensities for uninfected cells (HSV-1 - cells) versus infected cells (HSV-1 + cells) within the same infected sample, as well as uninfected cells versus infected cells within the same ACV-treated sample. (E) Concentrations of Aβ42/40/38 in pg/mL, Aβ42/40 ratios and Aβ42/38 ratios detected from conditioned media that had undergone heat inactivation for uninfected (control) dcOrgs, HSV-1 infected (HSV-1+) dcOrgs, HSV-1 infected and ACV-treated (Treated) dcOrgs, and dcOrgs with UV-inactivated HSV-1 (UV-HSV-1). P -values shown were calculated using 1-sided Wilcoxon ranked sum test with comparison to control uninfected dcOrgs. (F) Concentrations of Aβ42/40/38 in pg/mL, β42/40 ratios and Aβ42/38 ratios detected from conditioned media that had undergone heat inactivation for uninfected (control) dcOrgs and IAV infected (IAV+) dcOrgs. P -values shown were calculated using 1-sided Wilcoxon ranked sum test with comparison to control uninfected dcOrgs.

Article Snippet: The antibodies used in our study were: Alexa Fluor 647-conjugated Aβ1-42 (Bioss Antibodies bs-0107R-BF647) at a 1:50 dilution, Alexa Fluor 647-conjugated Tau (Thr212) (Bioss Antibodies bs-5420R-BF647) at a 1:50 dilution, Alexa Fluor 647-conjugated Solanezumab (Novus Biologicals FAB9919R) at a 1:50 dilution, Alexa Fluor 647-conjugated TRA-1-60 (BioLegend 330605) at a 1:20 dilution, Alexa Fluor 647-conjugated Nestin (Novus Biologicals IC1259R-100UG) at a 1:50 dilution, Alexa Fluor 647-conjugated EOMES (Novus Biologicals IC6166R-100UG) at a 1:50 dilution, APC-conjugated TuJ1 (Biolegend 801219) at a 1:20 dilution, Alexa Fluor 647-conjugated NeuN (Novus Biologicals NBP1-92693AF647) at a 1:50 dilution, Alexa Fluor 647-conjugated VMAT2 (R&D Systems FAB8327R) at a 1:20 dilution, Alexa Fluor 647-conjugated GFAP (BioLegend 644706) at a 1:20 dilution, APC-conjugated GLAST (Miltenyi 130-123-555) at a 1:50 dilution, Alexa Fluor 647-conjugated Iba1 (Novus Biologicals 603102) at a 1:50 dilution, APC-conjugated P2RY12 (BioLegend 392113) at a 1:20 dilution, Alexa Fluor 647-conjugated CD4 (Biolegend 300520) at a 1:20 dilution, Alexa Fluor 647-conjugated Olig1/2/3 (Bio-techne FAB2230R-MTO) at a 1:50 dilution, Alexa Fluor 647-conjugated O4 (R&D Systems FAB1326R) at a 1:20 dilution and Alexa Fluor 647-conjugated O1 (R&D Systems FAB1327R) at a 1:20 dilution.

Techniques: Infection, Control, Comparison

Journal: Neurotherapeutics

Article Title: Discovery of an APP-selective BACE1 inhibitor for Alzheimer's disease

doi: 10.1016/j.neurot.2025.e00610

Figure Lengend Snippet: Inhibitory activity and brain permeability of FAH65E(-) . Shown are (A) dose-response curves for FAH65 racemate and the FAH65E(+) and (-) enantiomers in the P5-P5′ assay and (B) sAPPβ and (C) Aβ1-42 in CHO-7W cells after treatment with increasing concentrations of FAH65 racemate and enantiomers. Legend in B also applies to C. Data graphed as the mean and SEM. (D) FAH65E(-) (black line) and sAPP β (blue dashed line) levels in brain from ApoE4TR-5XFAD mice after oral delivery of 30 ​mg/kg FAH65E(-) doses are shown. PK-PD study design did not include 0 h timepoint untreated mice but included time points 1, 2, 4 and 6 ​h after last dose on Day 2. N ​= ​3 mice per time point.

Article Snippet: Media was assayed using an AlphaLISA for Aβ1-42 (Perkin Elmer catalog # AL276C), sAPPα (R&D Systems catalog # AF1168 conjugated with Perkin Elmer acceptor beads catalog # 6772001 + 2B3 antibody from IBL catalog # 11088 biotinylated), and sAPPβ (Perkin Elmer AL276-acceptor + IBL catalog # 18957 biotinylated).

Techniques: Activity Assay, Permeability

Journal: Journal of Biological Chemistry

Article Title: Inhibition of Apoptosis by Amphiregulin via an Insulin-like Growth Factor-1 Receptor-dependent Pathway in Non-small Cell Lung Cancer Cell Lines

doi: 10.1074/jbc.m207584200

Figure Lengend Snippet: FIG. 5. Effect of H358-conditioned medium, AR, and IGF1 on IGF1-R tyrosine phosphorylation. Serum-deprived H322 cells were incubated: A, for 30 min in serum-free medium (), in serum-free medium with IGF1 50 ng/ml (IGF1), or for indicated times in serum-free H358 CM (CM); B, for 30 min in serum-free medium (), in H358 CM (CM), in H358 CM preincubated with anti-AR neutralizing antibody (CM Ac AR), or in serum-free medium with 50 ng/ml of IGF1 (IGF1) or AR (AR), prior to detergent lysis and immunoprecipitation of the IGF1-R subunit. C, serum-deprived H358 cells were incubated for 30 min in serum-free medium (), in H358 CM (CM), or in serum-free medium with 50 ng/ml of IGF1 (IGF1) or AR (AR), prior to detergent lysis and immunoprecipitation of the IGF1-R subunit. Tyrosine phos- phorylation was visualized by protein immunoblotting with monoclonal anti-phosphotyrosine IgG as described (upper lane). Equal loading of immunoprecipitated proteins was confirmed by reprobing immunoblots with polyclonal anti-IGF1-R subunit antibody (lower lane). IgG, im- munoglobulin control for immunoprecipitation. Data are representative of at least three separate experiments.

Article Snippet: Primary monoclonal anti-human AR antibody (2 g/ml, R&D Systems) was coated to the bottom of 96-well plates and incubated overnight at room temperature.

Techniques: Phospho-proteomics, Incubation, Lysis, Immunoprecipitation, Western Blot, Control