Mem per plus

Integral membrane proteins and membrane-associated proteins from LNCaP cells were extracted by using Mem-PER Plus membrane protein extraction kit (Thermo Fisher Scientific, Waltham, MA, USA), according to the manufacturer’s instructions. One hundred micrograms of membrane-protein extracts and 40 μg of proteins from whole cell lysates were prepared in Laemmli sample buffer and resolved by 12% SDS-PAGE prior to immunoblotting with anti-Cx43 and anti-Cadherin11, as previously described in Section 4.5.

After 18 h of culture in a transwell assay, mMSCs and supernatants were collected and centrifuged to remove debris. Then, mMSCs were lysed by Membrane Protein Extraction Kit (Mem-PER™ Plus, Thermo Scientific, USA). CXCL12 (SDF-1α), vascular cell adhesion molecule-1 (VCAM-1), Clusters of Differentiation-44 (CD-44), matrix metalloproteinase2 (MMP2), Collagen-I, tumour necrosis factor (TNF-α) and IL-10 were measured by enzyme-linked immunosorbent assay (ELISA) kits (RayBiotech, USA; Bio-Techne, USA; Cusabio Biotech, China; ExCell Biology, China). The content of CXCL12 was determined using a semi-quantitative standard curve, and the additional CXCL12 was removed. ELISA was performed according to the manufacturer’s instructions. All samples were measured three times.