Tandem mass tag isobaric reagents

Tandem mass tag (TMT) isobaric reagents were from Thermo-Fisher Scientific (Waltham, MA). Water and organic solvents were from J.T. Baker (Center Valley, PA). Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) were from LifeTechnologies (Waltham, MA). Unless otherwise noted, all other chemicals, were from Sigma (St. Louis, MO).

Tandem Mass Tag (TMT) isobaric reagents, BCA Protein Concentration Kit, Protease inhibitor tablets, and trypsin were purchased from ThermoFisher Scientific (Rockford, IL). StageTip Empore-C18 material was purchased from CDS (Oxford, PA). Sep-Pak cartridges (50 mg) were from Waters (Milford, MA). LysC protease was from Fujifilm Wako (Richmond, VA). Mass spectrometry-grade water and organic solvents were from J.T. Baker (Center Valley, PA). The S. cerevisiae strain used was BY4716 from ThermoFisher Scientific (Waltham, MA). The S. pombe strain was NRRL Y-164 purchased from ATCC (Manassas, Virginia). The E. coli strain was purchased from Carolina Biological Supply Co. (Burlington, NC). Synthetic complete (SC), Edinburgh Minimal Media (EMM), yeast extracts with supplements (YES), and yeast-peptone-dextrose (YPD) media was from Sunrise Science (San Diego, CA). Luria-Bertani broth (LB) and Nutrient broth #1 (NB) was purchased from Millipore-Sigma (St. Louis, MO). Media formulations are available in the Supplemental Methods. Sera-MagSpeed Beads (cat. nos. 45152105050350 and 65152105050350) were from GE Life Sciences (Marlborough, MA).

The tandem mass tag (TMT) isobaric reagents and the S. cerevisiae strain used (BY4716) were from ThermoFisher Scientific (Waltham, MA). Water and organic solvents were from J.T. Baker (Center Valley, PA). The Dulbecco’s modified eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) was from LifeTechnologies (Waltham, MA). Trypsin was purchased from Pierce Biotechnology (Rockford, IL), and LysC was from Wako Chemicals (Richmond, VA). The HCT116 cell line (CCL-247) was purchased from ATCC (Manassas, VA). Unless otherwise noted, all other chemicals were from Pierce Biotechnology (Rockford, IL).

Tandem mass tag (TMT) isobaric reagents were from Thermo Fisher Scientific (Waltham, MA). Nicotine was purchased from Sigma (St. Louis, MO). Water and organic solvents were from J.T. Baker (Center Valley, PA). Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) were from LifeTechnologies (Waltham, MA). Trypsin was purchased from Pierce Biotechnology (Rockford, IL) and Lys-C from Wako Chemicals (Richmond, VA). Unless otherwise noted, all other chemicals were from Pierce Biotechnology (Rockford, IL). The HAP1wt cell line (C859) and the CRISPR/Cas edited α7KO cell line (HZGHC003352c012) were from Horizon Discovery (Saint Louis, MO).

Tandem mass tag (TMT) isobaric reagents were from ThermoFisher Scientific (Waltham, MA). Water and organic solvents were from J.T. Baker (Center Valley, PA). Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) were from LifeTechnologies (Waltham, MA). Unless otherwise noted, all other chemicals were from Sigma (St. Louis, MO). The antibodies used in this experiment were purchased from SantaCruz Biotechnology (Dallas, Texas): α1 (sc-65829), α2 (sc-365251), α3 (sc-365479), α4 (sc-1772), α5 (sc-376979), α6 (sc-376966), α7 (sc-5544), α9 (sc-13806) or Cell Signaling Technology (Beverley, MA): actin (4968). The human PaSC cell line used in this experiment was RLT-PSC (24 (link)).

Tandem Mass Tag (TMT) isobaric reagents, BCA Protein Concentration Kit, Protease inhibitor tablets, and trypsin were purchased from ThermoFisher Scientific (Rockford, IL). StageTip Empore-C18 material was purchased from 3M (St. Paul, MN). Sep-Pak cartridges (50 mg) were from Waters (Milford, MA). Lys-C protease was from Fujifilm Wako (Richmond, VA). Mass spectrometry-grade water and organic solvents were from J.T. Baker (Center Valley, PA). The S. cerevisiae strain used was BY4716 from ThermoFisher Scientific (Waltham, MA). The S. pombe strain was NRRL Y-164 purchased from ATCC (Manassas, Virginia). Synthetic complete (SC) and Edinburgh Minimal Media (EMM) media was from Sunrise Science (San Diego, CA). Glucose and pyruvate were from MilliporeSigma (St. Louis, MO).

Dulbecco's modified Eagle's-F12 medium (DMEM/F12; 11330) was purchased from Gibco (Carlsbad, CA). Fetal bovine serum (FBS; F0392) was purchased from Sigma (St. Louis, MO). CellStripper (25-056-CL) for non-enzymatic cell dislodgement was purchased from Mediatech (Manassas, VA). Tandem mass tag (TMT) isobaric reagents were from Thermo Scientific (Rockford, IL). Water and organic solvents were purchased from J.T. Baker (Center Valley, PA). (−)-Nicotine (≥99%) (N3876) was purchased from Sigma (St. Louis, MO). Sequencing-grade modified trypsin (V5111) was obtained from Promega (Madison, WI). Unless otherwise noted, other reagents and solvents were from Sigma-Aldrich and Burdick & Jackson, respectively. Primary antibody against amyloid precursor protein (ab15272) was from Abcam (Cambridge, MA), while secondary horseradish peroxidase anti-rabbit antibody (sc-2313) was from SantaCruz Biotechnology (Santa Cruz, CA).