Anti fus tls

Cultured neurons were harvested at 3 weeks of differentiation as cell pellets. Protein samples were prepared and assayed by the method described previously [23 (link)]. The following primary antibodies were used—anti-TDP43 (1:200, Proteintech), anti-FUS/TLS (1:200, Santa Cruz Biotechnology), Tau5 anti-tau (1:1000, Thermo Fisher Scientific), AT8 anti-p-tau (1:1000, Thermo Fisher Scientific), and anti-caspase-3 (1:200, Millipore).

PTE, green tea extract (GTE), and black tea extract (BTE) were kindly supplied by the China Academy of Pu-Erh Tea Research [21 (link), 22 (link)]. These extracts were dissolved in distilled water and adjusted to pH 7.4 with 1 M NaOH. Ammonium chloride (NH₄Cl), a lysosomal inhibitor, was purchased from Wako Pure Chemical Industries (Osaka, Japan). Lactacystin (Calbiochem, Bad Soden, Germany) was dissolved in dimethyl sulfoxide (DMSO). Sodium arsenite solution was purchased from EMD Millipore (Darmstadt, Germany). Anti-FUS/TLS, anti-LDLR (low density lipoprotein receptor), anti-TDP-43, anti-V5, anti-β-actin, and horseradish peroxidase-conjugated secondary antibody were purchased from Santa Cruz Biotechnology (Santa Cruz, CA), Abnova (Taipei City, Taiwan), Cell Signaling Technology (Beverley, MA), Invitrogen (Carlsbad, CA), Sigma (St. Louis, MO), and Thermo Scientific (Waltham, MA), respectively. Anti-EWS and anti-TAF15 were purchased from Cell Signaling Technology (Beverley, MA). The SOD1 antibody was a kind gift from Dr. Noriko Fujiwara.

All neurons were prepared by the methods described in a previous report [23 (link)]. The following primary antibodies were used: anti-OCT4 (1:200, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-SOX2 (1:200, Millipore, Burlington, MA, USA), anti-NANOG (1:200, R&D Systems), anti-SSEA-4 (1:100, Developmental Studies Hybridoma Bank, Iowa City, IA, USA), anti-TRA-1-81 (1:100, Thermo Fisher Scientific), Tuj1 anti-tubulin beta III isoform (1:200, Millipore), Tuj1 anti-tubulin beta III (1:200, Abcam, Cambridge, UK), anti-SMA (1:100, DAKO, Santa Clara, CA, USA), anti-AFP (1:100, DAKO), anti-Nestin (1:200, R&D Systems), anti-Musashi (1:200, Millipore), anti-MAP2 (1:200,Millipore), anti-TDP43 (1:200, Proteintech, Rosemont, IL, USA), anti-FUS/TLS (1:200, Santa Cruz Biotechnology), p-Tau (AT8), and anti-Caspase-3 (1:100, Millipore).