Anti α6

The human prostate cancer cell lines C4-2, 22RV1 and DU145 were purchased from the Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences (Shanghai, China). The PEP06 and 33 polypeptide were synthesized by Ningbo Kangbei Biochemical Co., Ltd. (Ningbo, China). RPMI 1640 medium was purchased from Thermo Fischer Scientific Inc.(Waltham, MA, USA) FBS, penicillin and streptomycin were purchased from HyClone Laboratories Inc. (Logan, UT, USA). Trypsin was purchased from Invitrogen (Carlsbad, CA, USA). Matrigel basement membrane matrix was purchased from Seebio Biotechnology Co., Ltd. (Shanghai, China). Dimethyl sulfoxide (DMSO) was purchased from VWR International Co. Transwell cell inserts were purchased from Wuxi NEST Biotechnology Co., Ltd. (Wuxi, China) Rabbit anti-αv, anti-β3, anti-α6, anti-β1, anti-MMP2, anti-E-cadherin, anti-β-catenin, anti-vimentin, anti-MMP9, anti-AKT and anti-P-AKT monoclonal antibodies were purchased from Abcam Trading Co., Ltd. (Shanghai, China).

Eight micrometre cryostat sections were rehydrated in phosphate-buffered saline for 10 min. Tissue sections were blocked with 5% normal goat serum and co-stained with anti-αSMA (Sigma, St Louis, MO, Cat# A2547, 1:200 dilutions) and anti-α₆ (Abcam, Cambridge, MA, Cat#14-0495, 1:300 dilutions) antibodies diluted in phosphate-buffered saline containing 1% goat serum, 0.3% Triton X-100 and 0.01% sodium azide according to manufacturer's instructions. Fluorochrome-conjugated secondary antibodies (SouthernBiotech, Birmingham, AL) were used according to the manufacturer's recommendation. Nuclei were stained with DAPI (Thermo Fisher Scientific, Waltham, MA). Fluorescent signals were detected using a confocal laser-scanning microscope Zeiss LSM710 confocal microscope equipped with a digital colour camera (Oberkochen, Germany). All fluorescent images were generated using sequential laser scanning with only the corresponding single-wavelength laser line, activated using acousto-optical tunable filters to avoid cross-detection of either one of the fluorescence channels.