Rm02835

Actin cytoskeleton staining was performed to observe the formation of pseudopodia. The PCa cell line was seeded in confocal dishes at 1 × 10⁵ cells/well. Confocal dishes were washed three times with PBS followed by fixation with 4% paraformaldehyde (PFA, YJ0002, YongJing bio, Guangzhou, China) for 15 min. After another three washes with PBS, rhodamine phalloidin (1:100, RM02835, Abclonal, Wuhan, China) was added to the dishes and incubated for 1 h at 37 °C in the dark. The nuclei were stained with DAPI (2 μg/ml, G1012, Servicebio, Wuhan, China), and all incubations were followed by 3 PBST washes. Before collecting the images under a confocal microscope (Zeiss LSM 800), the cells were sealed with anti-fluorescence attenuation sealant (S2100, Solarbio, Beijing, China).

Primary antibodies against pDDR1-Y792(11994S; CST; 1:1000), DDR1 (10536-1-AP; Proteintech; 1:1000), YAP1 (13584-1-AP; Proteintech; 1:1000), pYAP1-S127 (AP0489; ABclonal; 1:1000), YWHAE (11648-2-AP; Proteintech; 1:1000) VE-cadherin (bs-4310R; Bioss; 1:1000) and GAPDH (BE0024; Easybio; 1:4000) were used for Western blotting. Rhodamine Phalloidin (RM02835; ABclonal; 1:200), Primary antibodies against YAP1 (66900-1-Ig; Proteintech; 1:200), DDR1 (10536-1-AP; Proteintech; 1:200), VE-Cadherin (sc-9989; Santa Cruz; 1:100), E-selectin (MA1-22165; Invitrogen; 1:200), ICAM1 (sc-7891; Santa Cruz; 1:100), VCAM1 (sc-13160; Santa Cruz; 1:100), Caveolin-1 (66067-1-Ig; Proteintech; 1:200), Rab 5A (sc-166600; Santa Cruz; 1:100), PE-conjugated CD63 (BC-353003; BaiCheng Technology; 1:200) and LATS1 (3477S; CST; 1:200) were used for immunofluorescence staining. DDR1 antibody (200 μg/ml; sc-390268; Santa Cruz) were used for bead pulling/magnetic tweezer system. The validation of all primary antibodies for the species and application were described on the manufacturer’s website.