Fig. 3: α-synuclein PFFs induce neurotoxic activity in midbrain astrocytes.
From: Fibrillar α-synuclein induces neurotoxic astrocyte activation via RIP kinase signaling and NF-κB
a–f Primary human midbrain astrocytes were treated with transcription factor (TF) inhibitors and/or PFFs, as indicated. After 24 h, an astrocyte conditioned medium (ACM) was applied (1:1) to differentiated SH-SY5Y cultures for 24 h followed by endpoint analyses. b, c Viability of SH-SY5Y cells following treatment with ACM derived from astrocyte cultures treated with the indicated inhibitors was measured via ATP-luciferase assay (Cell Titer Glo). d, e Cell death of SH-SY5Y cells following 24 h ACM treatment was detected via TUNEL (d) and quantified as a percentage of TUNEL+ nuclei (e). f Levels of Caspase 3/7 activity in SH-SY5Y cells following treatments as in (a) was measured using a chromogenic DEVD-cleavage assay. g Modification of experimental setup in (a), in which SH-SY5Y cultures were pretreated with programmed cell death (PCD) inhibitors for 30 min prior to addition of ACM. h Viability of SH-SY5Y cells following treatment with ACM and PCD inhibitors as in (g) was measured via ATP-luciferase assay (Cell Titer Glo). ns not significant, *p < 0.05, **p < 0.01, ***p < 0.001. Bars represent group means. n = 6 independent replicates for all experiments.
