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. 2014 Jul 24;9(7):e102186. doi: 10.1371/journal.pone.0102186

Figure 4. CDC42 signaling regulates chronic inflammation associated with senescence.

Figure 4

(A) Immunostaining for p53, p21, and RELA in sections of the lungs and the renal glomeruli from endothelial cell-specific Mdm2 conditional knockout mice (Mdm2 CKO, Pdgfb-Cre-ER; Mdm2 loxP/loxP) and their littermate controls (Cont, Mdm2 loxP/loxP). Arrowheads indicate positive staining of capillary endothelial cells. Scale bar = 100 µm. (B) Expression of p21 and pro-inflammatory genes in the lungs of Mdm2 CKO mice (Pdgfb-Cre-ER; Mdm2 loxP/loxP), their littermate controls (Cont, Mdm2 loxP/loxP), Cdc42 CKO mice (Pdgfb-Cre-ER; Cdc42 loxP/loxP), and Mdm2 & Cdc42 CKO mice (Pdgfb-Cre-ER; Mdm2 loxP/loxP; Cdc42 loxP/loxP) was examined by real-time PCR. n = 4–6. (C) Expression of Cdc42 was examined by real-time PCR in the lungs of Cdc42 CKO mice (Pdgfb-Cre-ER; Cdc42 loxP/loxP), their littermate controls (Cont 1, Cdc42 loxP/loxP), Mdm2 & Cdc42 CKO mice (Pdgfb-Cre-ER; Mdm2 loxP/loxP; Cdc42 loxP/loxP), and their littermate controls (Cont 2, Mdm2 loxP/loxP; Cdc42 loxP/loxP). n = 5–6. (D) Immunostaining for p21 in paraffin-embedded sections of the aorta from Apoe knockout mice (Apoe KO, Apoe −/−) and wild-type littermates (WT, Apoe +/+). Black arrowheads indicate negative staining of aortic endothelial cells. Red arrowheads indicate positive staining. Scale bar = 100 µm. (E) Expression of Cdkn1a (p21), Cd68, and pro-inflammatory genes in the aortas of Cdc42 CKO mice (Pdgfb-Cre-ER; Cdc42 loxP/loxP), their littermate controls (Cont, Cdc42 loxP/loxP), Apoe KO mice (Apoe −/−;Cdc42 loxP/loxP), and Apoe KO & Cdc42 CKO mice (Apoe −/−; Pdgfb-Cre-ER; Cdc42 loxP/loxP) was examined by real-time PCR. n = 4–6. (F) Immunostaining for p21 in frozen sections of the aorta from Apoe KO mice (Apoe −/−), wild-type littermates (WT, Apoe +/+), Apoe KO littermates (Apoe −/−;Cdc42 loxP/loxP), and Apoe KO & Cdc42 CKO mice (Apoe −/−; Pdgfb-Cre-ER; Cdc42 loxP/loxP). Black arrowheads indicate negative staining of aortic endothelial cells for p21. Red arrowheads indicate positive staining for p21. Scale bar = 20 µm. (G) Oil red O staining of aortic sinus sections from Apoe KO mice (Apoe −/−;Cdc42 loxP/loxP) and Apoe KO & Cdc42 CKO mice (Apoe −/−; Pdgfb-Cre-ER; Cdc42 loxP/loxP). Scale bar = 100 µm. (H) Quantification of the atherosclerotic lesion area relative to the total area at the level of the aortic sinus in Apoe KO mice and Apoe KO & Cdc42 CKO mice. n = 5. Data are shown as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001.