Anti rab11a

Dissected mammary fat pads were fixed in MethaCarn and embedded in paraffin. Seven μm-thick sections were deparaffinized before staining with primary antibodies (overnight, 4°C), and secondary antibodies (1 h, room temperature). Nuclei were counterstained with DAPI. Primary antibodies used were: rabbit polyclonal anti-PAR3 (1:200; Chemicon), anti-aPKC (1:200; clone C-20, Santa Cruz Biotechnology), anti-RAB11A (1:200; Abcam), anti-pSTAT5 (Tyr694, 1:100; Cell Signalling), anti-cleaved caspase 3 (1:100; Cell Signalling), anti-WAP (1:300; clone R-131, Santa Cruz Biotechnology) and anti-keratin 5 (K5) (1:2,000; Covance); rabbit monoclonal anti-KI67 (1:100; clone SP6, Neo Markers); and mouse monoclonal anti-HTT (1:300; 4C8), anti-E-cadherin (1:200; BD Bioscience) and anti-GM130 (1:100; BD Bioscience). Antigen retrieval was performed by boiling the slides for 10 min in a microwave in 10 mM citrate buffer (pH 6) for cleaved caspase 3, Ki67, WAP, and p-STAT5A, or in EDTA buffer (pH 8.8) for 10 min for PAR3, aPKC, RAB11A, HTT, GM130, K5, and E-cadherin antibodies. Secondary antibodies used were goat anti-mouse and anti-rabbit conjugated to AlexaFluor-488 or AlexaFluor-555 or Biotin (Vector Laboratories).

Hoechst 33342 and Lyso-Tracker Red were purchased from Beyotime Biotechnology (Shanghai, China); Lipofectamine 2000 from Thermo Fisher Scientific (Shanghai, China); Bafilomycin A1 and Torin1 from Selleck Chemicals (Houston, USA); Puromycin from MedChemExpress (New Jersey, USA); Adenovirus Ad-mRFP-GFP-LC3 and Ad-GFP-LC3 from HANBIO (Shanghai, China); polybrene from Sigma-Aldrich (Saint Louis, USA); Anti-Actin, Anti-LC3, Anti-ATG5, Anti-Rab11A, and Anti-Rab18 monoclonal antibodies from Abcam (Shanghai, China); Anti-β-Tubulin polyclonal antibodies, goat anti-mouse and goat anti-rabbit HRP-conjugated Ig G (H + L) from FdBio Science (Hangzhou, China); ChamQ SYBR qPCR Master Mix (High ROX Premixed) from Vazyme (Nanjing, China); Protease inhibitor cocktail (Bimake, Texas, USA); TRIzol reagent and Goat anti-Rabbit IgG (H + L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ Plus 594, from Invitrogen (Shanghai, China); ReverTra Ace qPCR RT Kit from TOYOBO (Oosaka, Japan).

The following primary antibodies were used; CD11a Alexa Fluor^™ 594 clone HI111 (Biolegend, San Diego, CA), rabbit polyclonal anti-Rab5, anti-Rab11A, anti-Rab7, or anti-Lamp1 antibody (Abcam, Cambridge, UK), anti-beta-actin antibody (AnaSpec, Fremont, CA) (1:1000), and anti-LtxA monoclonal antibody 107A3A3 [75 (link)] in hybridoma supernatants (1:10 dilution). The following secondary antibodies were used: goat anti-rabbit IgG Alexa Fluor^®488 (1:1000); horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (Fc) or (HRP)-goat anti-rabbit (Pierce, Rockford, IL) (1:10,000). Transferrin labeled with Alexa Fluor®555 was from Invitrogen (Waltham, MA, USA). Dynamin inhibitor Dynole 34-2 and its inactive control, Dynole 31-2, were purchased from SigmaAldrich (St. Louis, MO), Dynasore and Pitstop 2 (Abcam, Cambridge, UK). The inhibitors were used in the following concentrations: 10 μM Dynole 34-2; 10 μM Dynole 31-2; 10 μM Dynasore; 5 μM Pitstop 2.