Fig 1: CRISPR/Cas13d regulated the function of GZMA through controlling the expression of miR-25-3p and PAR1 in PARs pathway in chronic apical periodontitis. A, B The PAR1, TGF-β mRNA and protein expression levels were decreased significantly after inhibition of GZMA via crRNA/Cas13d. C, D The GZMA mRNA and protein expression levels were no changed after knockdown of TGF-β. E The miR-25-3p expression was promoted significantly after inhibition of GZMA. F, G After transfected with miR-25-3p inhibitor, the GZMA mRNA and protein expression levels were no changed. While, the TGF-β mRNA and protein expression levels were increased dramatically. H The expression of XPO5 was increased significantly. I The GZMA protein expression level was no changed after transfection of XPO5 siRNA. *Means P < 0.05, **means P < 0.01, ***means P < 0.001
Fig 2: The schematic diagram of GZMA in osteoclasts. GZMA degrades exportin 5 that transports miR-25-3p pre-miRNA from the nucleus to the cytoplasm of osteoclast, and suppresses the expression of miR-25-3p. TGF-β promotes the expression of PAR1 and finally facilitates cell growth, inhibited cell apoptosis in osteoclasts
Fig 3: Molecular docking analysis to assess the interaction between F2R and dasatinib and an evaluation of dasatinib’s impact on MCF-7 cells. (a)–(d) Top four molecular docking sites between F2R and dasatinib. (e) Comparison of F2R mRNA expression levels in MCF-12A and MCF-7 cells. (f)–(j) The MTT assay was employed to determine the IC50 of dasatinib in MCF-7 cells and assess cell viability following treatment with 6 and 8 μM/L dasatinib. Additionally, the cellular state of MCF-7 cells treated with 6 μM/L dasatinib for 48 h was observed, and a scratch assay was conducted to evaluate the migration ability of MCF-7 cells treated with the same concentration of dasatinib for 48 h. Furthermore, the levels of F2R protein in MCF-7 cells after treatment with 6 μM/L dasatinib for 48 h were measured using the F2R ELISA Kit. **p < 0.01, ***p < 0.001, ###p < 0.001.
Fig 4: Predicted drug responsiveness in low and high F2R expression groups. (a)–(l) Exemplary drugs displaying heightened sensitivity in high-F2R expression groups, encompassing sunitinib, crizotinib, pazopanib, TAE684, AC220, saracatinib, PD-0325901, MG-132, temsirolimus, dasatinib, WH-4-023, and TGX221.
Fig 5: Gene enrichment analysis to identify pathways associated with F2R. (a) Genes highly correlated with F2R (correlation coefficient >0.6) were identified in BC using Pearson’s correlation analysis. (b) A subset of 76 DEGs exhibiting a strong correlation with F2R were selected for further investigation. (c) Gene ontology (GO) enrichment analysis was performed to explore the functional characteristics of F2R-related DEGs. (d) KEGG enrichment analysis was conducted to identify pathways associated with F2R-related DEGs. (e) GSEA was utilized to elucidate the signal pathways associated with F2R mRNA expression in BC.
Supplier Page from Thermo Fisher Scientific for Human PAR1 ELISA Kit