Culture slides

To determine the cellular localization of LINC00885, FISH was performed by utilizing the RNAscope Multiplex Fluorescent Reagent Kit v2 (Advanced Cell Diagnostics, California, USA). CC cells were subjected to 24-h incubation on the culture slides (Thermo Fisher Scientific, Inc., Waltham, MA, USA), followed by 0.5-h fixation in 4% paraformaldehyde. Then, the cell slides were treated with Hydrogen Peroxide and Protease III, followed by 2-h hybridization with the LINC00885 FISH probe (RiboBio, Guangzhou, China) in the HybEZ Oven (Advanced Cell Diagnostics). Thereafter, cells were washed and subjected to dehydration in the graded ethanol. Finally, slides were stained with DAPI (Cell Signaling Technologies, Danvers, USA). The observation and photographing of the cell slides was accomplished by using a FV1000 confocal laser microscope (Olympus, Tokyo, Japan).

Human brain cells and postmortem brain tissues on culture slides (Thermo Scientific, Waltham, MA, USA) were fixed in 4% paraformaldehyde, permeabilized with 0.1% Triton X-100, blocked in 10% milk/0.1% goat serum, and immunolabeled. Primary antibodies from Thermo Fisher Scientific (Waltham, MA, USA) included Occludin conjugated to the Alexa Fluor 488 dye at 1:1000 dilution (catalog # 331588), from Abcam (Boston, MA, USA) included LRP-1 at 1:100 dilution (catalog # ab-92544), from Millipore (Burlington, MA, USA) included GFAP at 1:400 dilution (catalog # MAB360), and MAP-2 at 1:100 dilution (catalog # MAB378), and from Wako chemicals (Richmond, VA, USA) included Iba-1 at 1:50 dilution (catalog # 019-19741). A secondary antibody from Molecular Probes (Carlsbad, CA, USA) was used to visualize immunoreactivity and 4′,6-diamidino-2-phenylindole (DAPI) from Thermo Fisher Scientific (Waltham, MA, USA) was used to label cell nuclei. Cells were imaged using an inverted fluorescence microscope (Zeiss, Germany).