Cr0013

Lung tissue paraffin sections were dewaxed according to the manufacturer’s instructions, incubated with 0.3% TritonX-100 (Sinopharm, 30188928, Shanghai, China) for 8 min, blocked with 10% goat serum (Solarbio, SL038, Beijing, China) for 50 min, then incubated with the antibody of α-SMA (1:200, Abcam, Ab7817, Cambridge, UK) at 4 °C overnight. On the second day, sections were rinsed with 1×PBS solution (Sparkjade, CR0013, Jinan, China) and incubated with anti-IgG Fluor (1:200, Affinity, S0006, Beijing, China) for 1 h. The nuclei were stained with DAPI (1:400, Sigma, D9542, St. Louis, MO, USA). After 1×PBS washing, tissue sections were sealed with neutral glycerin and photos taken in an automatic living cell fluorescence microscopic imaging system (Invitrogen, EVOS M5000, Carlsbad, CA, USA).

lncITPF, U6, and 18S FISH probes were synthesized by Guangzhou RiboBio Co., Ltd. The experiment was performed with the Ribo lncRNA FISH Probe Mix according to the manufacturer’s protocol (RiboBio, C10910, Guangzhou, China). Cells were inoculated on a circular slide. When the cell density reached 50–60%, 4% paraformaldehyde (Meilunbio, MA0192, Dalian, China) was added. Then, cell samples were washed with 1 × PBS (Sparkjade, CR0013, Jinan, China) and punched with 0.3% TritonX-100 (Sinopharm, 30188928, Shanghai, China) at 4 °C for 3 min. Permeate solution was washed away with 1 × PBS and 200 μL pre-hybridization solution was added for 30 min. Subsequently, lncITPF, U6, and 18S FISH Probe Mix were added to the cell samples in a hybridization oven at 37 °C overnight. On the second day, the hybridization probe solution was aspirated at 42 °C and washed with SSC (Solarbio, S1030, Beijing, China). Then DAPI (Sigma, D9542, St. Louis, MO, USA) solution was added for 6 min. Finally, fluorescence was observed using a laser confocal microscope.