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. 2021 Jul 31;12(8):756. doi: 10.1038/s41419-021-04049-0

Fig. 6. RIPK signaling is required for NF-κB activation downstream of α-synuclein PFFs.

Fig. 6

a Confocal microscopy of NF-κB component p65 (red) and nuclei (DAPI, blue) in human midbrain astrocytes following 2 h PFF treatment ± cotreatment with indicated inhibitors. Scale bars = 50 μm. b Colocalization of red and blue signal in (a) is quantified as Fisher’s Z transformed index of correlation. c Nuclear fractions were isolated from human midbrain astrocyte cultures following 2 h PFF treatment ± cotreatment with indicated inhibitors. Relative levels of p65 in nuclear fractions were quantified using ELISA. d–j Primary human midbrain astrocyte cultures were treated with PFFs or PBS control solution. Cultures were pretreated (30 min) with inhibitors of RIPK3 (GSK872) or NF-κB (BAY) signaling prior to the addition of PFFs. Levels of indicated transcripts were measured at indicated time points using qRT-PCR. ns not significant, *p < 0.05, **p < 0.01, ***p < 0.001. Bars represent group means. n = 6 independent replicates in (a–c). n = 3 independent replicates in (d–j).