Fig 1: TP and Ki20227 downregulate CSF1R protein expression. (a) Western blot images of CSF1R and GADPH protein expressions of experimental groups. (b) Quantitative analyses of CSF1R protein expression. Data were expressed as mean ± SEM. One-way ANOVA with Tukey's tests, n = 7. The WT vs. WT+stroke groups, ∗∗∗P < 0.001; the WT+Stroke+TP and WT+K+stroke groups vs. WT+stroke group, ###P < 0.001; the WT+stroke+K+TP group vs. WT+stroke+TP and WT+stroke+K groups, ^^^P < 0.001.
Fig 2: The mechanism of SMS2 inhibitors in inhibiting PANC-CM-induced M2 polarization. (A). Gene expression of the cytokine that can induce M2 polarization in PANC-1 (a) was analyzed by CCLE database and was detected by Realtime PCR in PANC-02 (b). The expression of MRC1 is positively correlated with CSF1R (c). The expression of CSF1R is positively correlated with SMS2 in pancreatic cancer (d). Immunoblotting was used to detect the expression of CSF1R and downstream proteins in BMDM (B) and THP-1 (C) after PDAC-CM stimulation for 48 h with or without YE2. Immunoblotting was used to detect the expression of CD206 and p-Stat3 in BMDM (E) and THP-1 (D) after rescue of p-Stat3 with Colivelin. Immunoblotting was used to detect the expression of CSF1R and M2 marker (Arg-1 or CD206) after supplement of exogenous sphingomyelin (SM) in BMDM (G) and THP-1 (F). The densitometry analysis data of BMDM (B,E and G) and THP-1 (C,D and F) was showed. *p < 0.05; **p < 0.01; ***p < 0.001.
Fig 3: Osteoclast differentiation process of the palatal bone (PPP). (A) TRAP staining from E14.5 to E18.5 “a–e”, von Kossa staining from E14.5 to E18.5 “f–j”. Immunofluorescence staining expression patterns of CD31 from E14.5 to E18.5 “k–o”. (B–E) Immunofluorescence staining expression patterns of CD14, F4/80, CSF1R and RANKL during palatal osteoclastogenesis from E14.5 to E17.5. Black triangles represent TRAP-positive regions, and white triangles represent mineralizing and CD31-positive regions. Scale bar 500 µm.
Fig 4: Osteoclast differentiation process of the palatal bone (PPMX-P). (A) TRAP staining from E14.5 to E18.5 “a–e”, von Kossa staining from E14.5 to E18.5 “f–j”. Immunofluorescence staining expression patterns of CD31 from E14.5 to E18.5 “k–o”. (B–E) Immunofluorescence staining expression patterns of CD14, F4/80, CSF1R and RANKL during palatal osteoclastogenesis from E14.5 to E17.5. Black triangles represent TRAP-positive regions, and white triangles represent mineralizing and CD31-positive regions. Scale bar 500 µm.
Fig 5: Osteoclast differentiation process of the palatal bone (PPMX-A). (A) TRAP staining from E14.5 to E18.5 “a–e”, von Kossa staining from E14.5 to E18.5 “f–j”. Immunofluorescence staining expression patterns of CD31 from E14.5 to E18.5 “k–o”. (B–E) Immunofluorescence staining expression patterns of CD14, F4/80, CSF1R and RANKL during palatal osteoclastogenesis from E14.5 to E17.5. Black triangles represent TRAP-positive regions, and white triangles represent mineralizing and CD31-positive regions. Scale bar 500 µm.
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