Sc 362281

AS1411 was labeled with FITC. 4T1 and B16F10 cells (5 × 10⁴ cells per well) were seeded into a glass-bottom cell culture dish (20 mm). When the cell density reached 70%, the cells were transfected with AS1411 (800 ng per well) using Lipo3000 (Invitrogen, USA) for 24 hours according to the instructions. The treated cells were fixed in 4% paraformaldehyde for 15 min at room temperature. Immunofluorescence was performed as described previously (58 (link)). Antibodies used were anti-nucleolin (1:100; Abcam, ab129200) and Dylight 594 goat anti-rabbit immunoglobulin G (1:500; sc-362281, Santa Cruz Biotechnology). The images and videos were taken using a confocal microscope (Zeiss LSM 880, Germany).

CRC tissues were fixed in 4% formaldehyde solution for 2 h at 25°C and then sectioned into 5-µM-thick frozen sections. The sections were washed in cold PBS 3 times and subsequently blocked with 2% bovine serum albumin V at 25°C (BSA-V; Beijing Solarbio Science & Technology Co., Ltd., Beijing, China) for 1 h. Samples were incubated with primary antibodies against E-cadherin (1:20; sc-8426; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) diluted with 1% BSA-V overnight at 4°C. Following 3 washes with PBS, the sections were incubated with tetramethylrhodamine conjugated goat anti-rabbit secondary antibody (1:1,000; sc-362281; Santa Cruz Biotechnology, Inc.) diluted with 1% BSA-V in the dark for 1 h at 25°C and washed in PBS again for 3 times. DAPI diluted with PBS was used to stain the nuclei at 25°C. Images at a magnification of ×40 were captured using an inverted fluorescence microscope (Nikon Corp., Tokyo, Japan).