Pre hybridization buffer

GenePharma designed the FISH probes of SOX21-AS1. Cells were fixed and washed, and then were subject to permeabilization with 0.5% Triton X-100. Then, pre-hybridization buffer (Sigma-Aldrich, USA) was added with SOX21-AS1 probe. DAPI solution was used to redye cells and the fluorescent signal was observed under the microscope. The experiment was subject to three independent repeats.

Design and synthesis of HOXC-AS1-FISH probe were accomplished by Invitrogen. BGC-823 or AGS cells were plated on culture slides, fixed in paraformaldehyde (PFA; Sigma-Aldrich, St. Louis, MO, USA), followed by the sealing with prehybridization buffer (Sigma-Aldrich). Hybridization mixture was added with FISH probe. Slides were washed in buffer adding saline sodium citrate (SSC; Sigma-Aldrich). Cell nuclei were stained by DAPI (Sigma-Aldrich). Cells were examined with Olympus fluorescence microscope (Olympus, Tokyo, Japan).

GenePharma was used to design the FISH probes of KCNQ1OT1. T24 and HT-1197 cells were immobilized for 10 min in 4% PFA and washed with PBS (Sigma-Aldrich) for three times. Subsequently, PBS containing with 0.5% Triton X-100 was applied for cell permeabilization. Following washing, pre-hybridization buffer (Sigma-Aldrich) was added into each well to incubate, which lasted for half an hour. The slide was hybridized with KCNQ1OT1 FISH probe, and then was washed three times with 2× Saline Sodium Citrate (SSC; Sigma-Aldrich) at 42 °C. Hoechst 33342 solution (Invitrogen) was used to stain cells. Finally, fluorescent signal was observed by using the microscope [20 (link)].