G9545

The following antibodies were used for this study: P2ry12 (1:500), a generous gift from Dr. Oleg Butovsky; CD68 (1:200), Abcam ab53444, RRID:AB_869007; 4G8 (20 μg/reaction), Biolegend^® RRID:AB_10175149; Anti‐CD11b (1:100), BD Biosciences Cat# 552850 RRID:AB_394491; anti‐CD45 (1:200), eBioscience Cat# 17‐0451‐82 162 RRID:AB_469392; anti‐Ly6C (1:400), eBioscience Cat# 12‐5932‐82 RRID:AB_10804510; anti‐Ly6G (1:100), Biolegend 163 Cat# 127605 RRID: AB_1236488; GAPDH (1:5,000), Sigma‐Aldrich G9545 RRID: AB 796208; β2AR (1:500), Abcam ab182136 (lot: GR302897‐20) RRID: AB not available.
Detailed methods regarding environmental enrichment setup, mouse brain tissue staining and microglial morphology analysis, microglia isolation and RNA analysis, and human brain extract preparation and analysis are reported by previous publications (Yang et al, 2013, 2017; Xu et al, 2016). Methods unique to this study are listed below:

The human cervical cancer cell line HeLa and human lung cancer cell line A549 were cultured in Dulbecco's Modified Eagle's Medium (Sigma-Aldrich D5671) supplemented with 10% FBS and 1% penicillin/streptomycin at 37°C and 5% CO₂. Anti-GAPDH antibody (Sigma-Aldrich G9545) and anti-histone H3 antibody (Abcam ab24834) were used for western blot analysis.

Yeast culture, UV cross-linking, cell lysis, 2C method and 2C-western blot experiments were done as previously described (Asencio et al. 2018 (link)). The following antibodies were used in western blot experiments: Anti-Pab1 1:4000 (Abcam, #ab189635), Anti-GAPDH 1:4000, a polyclonal antibody that detects all three Tdh (Tdh1, Tdh2, and Tdh3) yeast GAPDH isoforms (Sigma-Aldrich #G9545 [33]), Anti-histone H3 HRP 1:1000 (Abcam, #ab21054), Anti-Hexokinase 1:10000 (Bio-Rad, #4959-9988), Anti-Tpi 1:4000 (Proteintech, #10713-1-AP), Anti-tubulin 1:4000 (Abcam, #ab6160), and Anti-PFK antibody 1:4000 (Heinisch 1986 (link)). Peroxidase anti-peroxidase (PAP) antibody 1:10,000 was used to detect all TAP and Protein A tagged proteins (Sigma-Aldrich # P1291); Anti-ATP5b 1:1000 (Proteintech, #17247-1-AP) was used to detect the yeast ortholog protein ATP1.