Anti actin

LPS, D-galactosamine were purchased from Sigma-Aldrich Co (St. Louis, MO, USA). AST, ALT, MPO kits were purchased from the Institute of Jiancheng Bioengineering (Nanjing, China). TNF-α, IL-1β, and IL-6 ELISA kits were purchased from Biolegend (San Diego, CA, USA). Gas6 was purchased from R&D Systems (Minneapolis, MN, USA). Antibodies used for western blot were as follows: anti-Bcl-2 (Proteintech, Rosemont, IL, USA), anti-Bax (Proteintech, Rosemont, IL, USA), anti-ß-actin (Proteintech, Rosemont, IL, USA), NF-κB signal pathway kit (Cell Signaling Technology, Beverly, MA, USA).

The proteins of rat stomach tissues were extracted in ice-cold RIPA lysis buffer (Solarbio, China) by ultrasound, and then determined by the enhanced bicinchoninic acid protein assay kit (Thermo, USA). 30 μg of each sample was loaded on 10% SDS-PAGE gels. And protein blots were transferred onto polyvinylidene fluoride membranes (Milipore, USA). After blocking with 5% non-fat milk, the blots were incubated overnight at 4 °C with a primary antibody: anti-actin (1:5,000, Proteintech Group) and anti-striatin (STRN, 1:1,000, Proteintech Group) antibodies. Then the membranes were washed with a mixture of Tris-buffered saline and Tween 20 (TBST) and incubated at room temperature for 1 h with a secondary antibody conjugated to horseradish peroxidase. Finally, the protein blots were visualized using an enhanced chemiluminescence kit (Millipore, USA).

The reagents used in this study include the following: cisplatin (Sigma‐Aldrich, St. Louis, MO), 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) (Sigma‐Aldrich) and ViaFect™ transfection reagent (Promega, Madison, MI). Antibodies used in this study include anti‐p62 (Abcam, Cambridge, MA, USA), anti‐LC3 (Abcam), anti‐Caspase 8 (Proteintech, Chicago, IL), anti‐actin (Proteintech) and anti‐ubiquitin (Santa Cruz, CA).

Primary antibodies: Anti-Atg5 (Cat# AP1812b) was purchased from Abgent, San Diego, USA. Anti-Atg6 (Cat# PD017) was from MBL, Nagoya, Japan. Anti-Atg16L1 (Cat# 8089) was purchased from Cell Signaling Technology, Pickering, Canada. Anti-GAPDH (glyceraldehyde-3-phosphate dehydrogenase, Cat# CW0100) was from CWBIO, Beijing, China. Anti-Actin (Cat# 14395-1-AP) was from Proteintech Group, Chicago, USA. Anti-Flag antibody (Cat# F3165) and monoclonal anti-LC3 (Cat# SAB4200361) were from Sigma-Aldrich, St Louis, USA. Anti-Myc (Cat# 11667149001) and anti-GFP (Cat# 11814460001) were from Roche Applied Science, Indianapolis, USA. Anti-Ppp1r36 was prepared by Beijing Huada Protein Innovation, Beijing, China.
Secondary antibodies: Goat anti-mouse IgG (H + L), horseradish peroxidase conjugated antibody (Cat# 31430) and goat anti-rabbit IgG (H + L), horseradish peroxidase conjugated antibody (Cat# 31460) were from Pierce Company, Rockford, USA. FITC-conjugated immunopure goat anti-rabbit IgG (H + L) (Cat# ZF-0311) was purchased from Feiyi Technology, Wuhan, China. Cy3-conjugated affinipure goat anti-rabbit IgG (H + L) (Cat# SA00009-2) was from Proteintech Group, Chicago, USA.

The human USP37 cDNA was subcloned into pcDNA3.1-Flag vector or PCDH vector and USP37-C350S was made using PCR-based site-directed mutagenesis method. All other constructs were generated using standard molecular cloning methods and were confirmed by DNA sequencing. Antibodies were commercially purchased: anti-USP37 (rabbit, 18465-1-AP, Proteintech), anti-Snail (rabbit, 3879, Cell Signaling), anti-Ubi (mouse, sc-8017, Santa Cruz), anti-actin (mouse, 60008-1-lg, Proteintech), anti-N-cadherin (mouse, 33-3900, Invitrogen), anti-HA (mouse, MMS-101P, Covance), anti-Flag (mouse, F3165, Sigma), anti-Flag (rabbit, F7425, Sigma), anti-Myc (mouse, 13-2500, Invitrogen), anti-GAPDH (mouse, 60004-1-Ig, Proteintech), normal IgG (rabbit, sc-2027, Santa Cruz), and GSH beads (GE). MG132 and cycloheximide (CHX) were obtained from Sigma.

Transgenic flies expressing the human TDP-43 (Wt or A315T-mutant) were described previously [40 (link),41 (link),87 (link)]. GMR-Gal4, OK371-Gal4, Elav-Gal4 and UAS-Lon-RNAi lines were obtained from the Bloomington Drosophila Stock Center (BDSC). Another UAS-Lon-RNAi fly line was obtained from the Vienna Drosophila Resource Center (VDRC). UAS-dLonOE was from the Kyoto Stock Center. The Tubulin-Gal80^ts (Tub-Gal80^ts) line was kindly provided by Dr. A. Guo (IBP, CAS) [48 (link)]. The UAS-mito-roGFP2-Grx1 fly lines were kindly provided by Dr. T. Dick [47 (link)].
For flies under the Elav-Gal4/Tub-Gal80^ts-driver or GMR-Gal4/Tub-Gal80^ts-driver, parental flies were crossed and cultured at 18°C, young flies after eclosion were transferred to 28°C for 4 hr every day to induce TDP-43 expression. Other flies were all cultured at 25°C. All flies were raised in standard fly food, 50% relative humidity, and 12hr-12hr light-dark cycles as described previously [41 (link),70 (link),87 (link)].
Antibodies used in this study include polyclonal rabbit-antibodies against TDP-43, ATP5A1, LonP1, HSPA9, ClpP, TOM20 and IMMT (ProteinTech Group Inc), as well as mouse monoclonal antibodies, anti-actin (ProteinTech Group Inc), anti-HSP60 (BD Biosciences) and anti-GAPDH (CWBIO). Rat-anti-dElav antibody is a kind gift from Dr. A. Guo.