Anti p62 sqstm1 sc 28359

CHX (01810), MG132 (C2211), CQ (C6628) and anti-Flag antibody (F3165) were obtained from Sigma (St. Louis, MO, USA). Anti-E-cadherin antibody (610181) was purchased from BD Biosciences (San Jose, CA, USA). Anti-vimentin (2870), N-cadherin (5741), HA (3724), Beclin 1 (3738), acetylated lysine (9441) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-p62/SQSTM1 (sc-28359) and GAPDH (sc-32233) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-LC3 (NB100–2220) antibody was obtained from NOVUS (Littleton, CO, USA).

Whole-cell lysate in radioimmunoprecipitation assay buffer, determination of protein concentration, SDS-PAGE, and immunoblotting were performed as described previously54 (link). For immunodetection, polyvinylidene fluoride membranes were blocked with 5% non-fat milk, incubated in Tris-buffered saline (pH 7.6; Sigma). The membranes were then incubated overnight at 4 °C with specific antibodies as follows: mouse monoclonal anti-AhR (sc-74571; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and anti-p62/SQSTM1 (sc-28359; Santa Cruz Biotechnology); rabbit monoclonal anti-LC3B (GTX127375; Genetex, Irvine, CA, USA), anti-ATG12-5 (GTX124181; Genetex), anti-ATG7 (GTX61647; Genetex), and anti-BNIP3 (ab109362; Abcam, Cambridge, UK), rabbit polyclonal anti-E-cadherin (GTX100443; Genetex) anti-vimentin (GTX100619; Genetex); and mouse monoclonal anti-β-actin (A1978; Sigma). The blots were then incubated with HRP-conjugated anti-rabbit and anti-mouse IgG antibodies obtained from Cell Signaling Technology (7074 and 7076; Danvers, MA, USA). Enhanced chemiluminescence detection was performed according to the manufacturer’s protocol (Millipore, Billerica, MA, USA).

The following antibodies were used for Western blot, immunofluorescence and PLA analyses: rabbit anti-BAG3 LP11 antibody (1:10000) was developed against a complete recombinant human BAG3 fused with glutathione S transferase (LP11; 1:10000 ) [16 (link)]; rabbit anti-HSPB8 against a C-terminal peptide (NELPQDSQEVTCT; 1:1000) [22 (link)]; anti-p62 SQSTM1 (sc-28359; 1:200) and anti-GFP (sc-9996, 1:1000; Santa Cruz); anti-GFP (632593, 1:500; Clontech); anti-HDAC6 (#7612; 1:1000) and anti-lysine-acetylated (ac-K-100, #9814, 1:1000; Cell Signaling Technologies); anti-vinculin (V9131; 1:5000), anti-Flag M2 (F3165, 1/200) and anti-α-tubulin mouse (T5168, 1:10000; Sigma-Aldrich); anti-cortactin (ab33333, 1:1000) and anti-α-tubulin rabbit (ab18251, 1:1000; Abcam); anti-acetyl-cortactin K309 (ABT1378, 1:1000); anti-cyclin B1 (ac-245, 1:1000) and anti-GAPDH Clone 6C5 (Fitzgerald, #10R-G109a, 1:10000) were from Millipore.
The Alexa-Fluor^® 488 Phalloidin (A12379, 1/50) comes from Molecular Probes/Thermo Fisher. Tubacin (S2239) was from Selleckchem; bisbenzimide H 33,342 (Hoechst, B2261), thymidine (T9250), fibronectin (F1141-5 mg) and CK666 (SML0006) were from Sigma-Aldrich.