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