Figure 7: CX3CR1 and VEGF are TGFβ1-targeting genes in mice and humans. | Nature Communications

Figure 7: CX3CR1 and VEGF are TGFβ1-targeting genes in mice and humans.

From: Activation of E-prostanoid 3 receptor in macrophages facilitates cardiac healing after myocardial infarction

Figure 7

(a,b) Schematic illustration of predicted Smad binding elements (SBEs) in the promoter of murine Cx3cr1 (a) and Vegf (b) genes. S=SBE. (c) Gel electrophoresis of PCR-amplified SBE-containing fragments in the promoter of the murine Cx3cr1 gene using anti-Smad2/3 immunoprecipitation. (d) Effect of the TGFβ1 blocker SB525334 on S3+S4 fragment-mediated luciferase activity. *P<0.05 as indicated (unpaired two-tailed t-test); n=4. (e) Effect of mutations of S3 (mS3) and S4 (mS4) on S3+S4 fragment-mediated transcription activity. *P<0.05 versus the S3+S4 group, n=4. (f) Effect of L-798,106 on S3+S4 fragment-mediated luciferase activity in RAW264.7 cells. *P<0.05 versus no treatment (unpaired two-tailed t-test);n=4. (g) Gel electrophoresis of PCR-amplified SBE-containing fragments in the promoter of the murine Vegf gene using anti-Smad2/3 immunoprecipitation. (h) Effect of SB525334 on the functional SBEs (S1′+S2′) in murine Vegf promoter. *P<0.05 as indicated (unpaired two-tailed t-test); n=4. (i) Effect of L798106 on S1′+S2′ fragment-mediated luciferase activity in RAW264.7 cells. *P<0.05 versus no treatment (unpaired two-tailed t-test); n=5. (j,k) Schematic illustration of the predicted SBE(s) on the human CX3CR1 gene (hS, (j)) and the human VEGF gene (hS′, (k)). (l) Gel electrophoresis of PCR-amplified SBE-containing fragments in the promoter of the human CX3CR1 gene. (m) Effect of SB525334 on hS-mediated transcription activity in THP-1 cells. *P<0.05 as indicated (unpaired two-tailed t-test); n=5. (n) Effect of L-798,106 on hS-mediated transcription activity in THP-1 cells. *P<0.05 versus no treatment (unpaired two-tailed t-test); n=5. (o) Gel electrophoresis of PCR-amplified SBE-containing fragments in the promoter of the human VEGF gene. (p) Effect of SB525334 on hS3′–hS5′ fragment-mediated transcription activity in THP-1 cells. *P<0.05 as indicated, n=5. (q) Effect of L-798,106 on hS3′–hS5′ fragment-mediated transcription activity in THP-1 cells. *P<0.05 versus no treatment (unpaired two-tailed t-test); n=5. (r) Schematic diagram of EP3-mediated regulation of CX3CR1 and VEGF genes in Mos/Mps through TGFβ1.

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