Anti pen2

Anti-PS1-N-terminus (1–65) (PRB-354P, 1:1,000, Covance, Davis, CA, USA); Anti-PS1-N-terminus (MAB1563, 1:100, EMD Millipore, Darmstadt, Germany); anti-PS1 loop (263–407) (529592, 1:2,000, Calbiochem); anti-APH1aL/C terminal (245–265) (PRB-550P, 1:1,000, Covance); anti-NCT (N1660, 1:1,000, Sigma), Anti-Pen2 (P5622, 1:1,000, Sigma); anti-BACE1 (AP7774b, 1:1,000, Abgent, Suzhou, China); anti-HA (H6908, 1:5,000, Sigma); anti-Flag (F3156, 1:2,000, Sigma); anti-A_2AR (05–717, 1:1,1000, Millipore); anti-EEA1 (610457, dilution 1:200, BD transduction laboratories).

Immunoblotting and immunostaining were performed with the following antibodies: anti-PS1 N (1–65) (529591, EMD anti-PS1, dilution 1:3 000, EMD Millipore, Darmstadt, Germany); anti-PS1 loop (263–407; 529592, dilution 1:2 000, Calbiochem); anti-Pen2 (P5622, dilution 1:1 000, Sigma); anti-NCT (N1660, dilution 1:1000, Sigma); anti-Aph1aL/C-terminal (245–265; PRB-550P, 1:1000, Covance, Davis, CA, USA); anti-BACE1 N-term (AP7774b, dilution 1:1000, Abgent, Suzhou, China); anti-APP-CTF (A8717, dilution 1:1000, Sigma); anti-Flag (F3156, dilution 1:2000, Sigma); anti-sAPPβ (6A1; 10321, dilution 1:1000, IBL, Hokkaido, Japan) anti-HA (H6908, dilution 1:5 000, Sigma); anti-c-Myc (9E10; sc-40, dilution 1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-NICD (D3B8; 4147, dilution 1:1000, Cell Signaling Technology, Beverly, MA, USA); anti-E-Cadherin-CTF (610182, dilution 1:1 000, BD Transduction Laboratories, San Jose, CA, USA); anti-APLP-1 C-Terminal (643–653; 171615, dilution 1:1000, Calbiochem); anti-Aβ 40 clone G2–10 (MABN11, dilution 1:1000, EMD Millipore); anti-Beta Amyloid, 1–16 (Monoclonal 6E10; SIG39300, dilution 1:1000, Covance); anti-calnexin (ab22595, 1:500, abcam, Cambridge, MA, USA); GM130 (610822, dilution 1:200, BD transduction laboratories); or EEA1 (610457, dilution 1:200, BD transduction laboratories); anti-GFAP (z0334, dilution 1:500, Dako, Aichi-ken, Japan).

Levodopa, bromocriptine, carbidopa, L-685,458, TAPI-1, Y-27632, and Forskolin (FSK) were purchased from Selleck Chemicals (Houston, TX, USA). Piribedil was from Tocris Bioscience (Bristol, UK). β-Secretase Inhibitor IV (BSI IV) was from Calbiochem (Hayward, CA, USA). Recombinant human BDNF, GNDF, IGF-I were from Peprotech (Rocky Hill, NJ, USA). cAMP and L-Ascorbic acid were from Sigma (St Louis, MO, USA). CellTiter-Glo was from Promega (Madison, WI, USA) and Effectene Transfection Reagent was purchased from QIAGEN (Hilden, Germany). Immunoblotting was performed with the following antibodies: anti-PS1 N-terminus (1–65) (PRB-354P, Covance, Davis, CA, USA); anti-APH1aL C-terminus (245–265) (PRB-550P, Covance); anti-NCT (N1660, Sigma); anti-Pen2 (P5622, Sigma); anti-BACE1 N-termimus (AP7774b, Abgent, Suzhou, China); anti-APP-CTF (A8717, Sigma); anti-sAPPα (secreted Amyloid Precursor Protein-α) (11088, IBL, Hokkaido, Japan); anti-HA (H6908, Sigma); anti-actin (A2066, Sigma); anti-D₂R (sc-9913, Santa Cruz Biotechnology, Santa Cruz, CA, USA). Immunostaining was performed with the following antibodies: anti-nestin (MAB353, Milipore, Darmstadt, Germany); anti-Sox2 (sc-17320, Santa Cruz); anti-Ki67 (556003, BD Transduction Laboratories, San Jose, CA, USA), and anti-Doublecortin (DCX) (sc-8066, Santa Cruz).