Additional file 1 of A missing piece of the puzzle in pulmonary fibrosis: anoikis resistance promotes fibroblast activation
Creators: name: Yin, Juan
name: Wang, Jing
name: Zhang, Xinxin
name: Liao, Yan
name: Luo, Wei
name: Wang, Sha
name: Ding, Jiawei
name: Huang, Jie
name: Chen, Mengling
name: Wang, Wei
name: Fang, Shencun
name: Chao, Jie
Publisher: name: figshare
Year: 2022
Subjects: subject: Biochemistry
subject: Medicine
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Physiology
subject: Pharmacology
subject: Biotechnology
subject: Ecology
subject: Immunology
subject: FOS: Clinical medicine
subject: Biological Sciences not elsewhere classified
subject: Information Systems not elsewhere classified
subject: Marine Biology
subject: Cancer
subject: Hematology
subject: Infectious Diseases
subject: FOS: Health sciences
subject: Virology

description: Additional file 1: Figure S1. (A) Statistical analysis of Masson’s trichrome staining of the fibrotic area. (n=5); *p &lt; 0.001 compared with the NS group. (B) Cell viability was detected using the CCK-8 assay after the three types of cells were treated with Con-CM and SiO2-CM for 6 h. Statistical analysis of cell viability. (n=5); *p &lt; 0.005 compared with the Con-CM group. (C) The expression levels of TrkB and Bcl2l11 were detected using Western blotting after HPF-a cells were treated with Con-CM and SiO2-CM for 24 h. Statistical analysis of cell viability (n=5); *p &lt; 0.05 compared with the Con-CM group. (D) The expression levels of TrkB and Bcl2l11 were detected using western blotting after BEAS-2B cells were treated with Con-CM and SiO2-CM for 24 h. Statistical analysis of cell viability (n=5); *p &lt; 0.05 compared with the Con-CM group. (E) The expression levels of TrkB and Bcl2l11 were detected using western blotting after HPMECs were treated with Con-CM and SiO2-CM for 24 h. Statistical analysis of cell viability (n=5); *p &lt; 0.05 compared with the Con-CM group. (F). Immunofluorescence staining showing the expression of the mesenchymal markers α-SMA and Col1a1 and the anoikis marker TrkB. Scale bar=200 μm. (H). Immunofluorescence staining showing the expression of the epithelial marker Cdh1 and the anoikis marker TrkB. Scale bar=100 μm. (I). Immunofluorescence staining showing the expression of the endothelial cell marker Cdh5 and the anoikis marker TrkB. Scale bar=100 μm. (J) Spatial transcriptomics showing the coexpression of Ntrk2 and Acta2. Figure S2. (A) Morphology of HPF-a cells after suspension culture for 48 h. Scale bar=100 μm. (B) BEAS-2B cells were suspended for 48 hours, cocultured with CM for 24 h, and double stained with Annexin V and PI. The sum of the counts in Q2-1, Q2-2 and Q2-4 was defined as the number of apoptotic cells. (C) HPMECs were treated as described in (B). The sum of the counts in Q2-1, Q2-2 and Q2-4 was defined as the number of apoptotic cells. (D) Statistical analysis of three independent experiments of BEAS-2B cell apoptosis assessed using flow cytometry (n=3); *p &lt; 0.05 compared with the attached group. (E). Statistical analysis of three independent experiments of HPMEC apoptosis assessed using flow cytometry (n=3); *p &lt; 0.05 compared the attached group. (F) Spatial transcriptomics showed the coexpression of Bim and Acta2. (G) Spatial transcriptomics showed the coexpression of Cas3 and Acta2. (H) Spatial transcriptomics showed the coexpression of Cas3 and Vimentin. (I) Spatial transcriptomics showed the coexpression of Cdk1 and Acta2. (J) Spatial transcriptomics showed the coexpression of Ccnb2 and Acta2. (K) Spatial transcriptomics showed the coexpression of Ccnb2 and Vimentin. Figure S3. (A) Representative western blot showing the transfection efficiency of ZC3H4-NICs in HPF-a cells. (B) Spatial transcriptomics showed the coexpression of Zc3h4 and Acta2. Figure S4. The expression of autophagy-related proteins (ATG5, BECN1 and LC3B) after CM stimulation of HPF-a cells. Figure S5. (A) Spatial transcriptomics showed the coexpression of Itga8 and Acta2. (B) Spatial transcriptomics showed the coexpression of Itgb1 and Vimentin.