Fig 2: Time-dependent fluctuation of CREB, aCaM kinase II, MAPK/Erk1/2 in PFC/FC nuclei. Modulation by chronic agomelatine. Expression levels of CREB (A), aCaM kinase II (C), MAPK/Erk1 (E) and MAPK/Erk2 (G) and phosphorylation levels of CREB at Ser133 (B), aCaM kinase II at Thr286 (D), MAPK/Erk1 at Thr202/204 (F) and MAPK/Erk2 at Thr202/204 (H) in nuclear fraction of PFC/FC from rats chronically treated with vehicle (VEH, black bar) or agomelatine (AGO, white bar) and sacrificed 1 or 16 h after the last injection. Data are expressed as % intensity units/mm2 (mean ± S.E.M.). Bonferroni post-hoc test (following 2-way ANOVA): *p < 0.05 VEH treated rats sacrificed 16 h after last injection vs. VEH treated rats sacrificed 1 h after last injection; § p < 0.05 AGO 16 h after last injection vs. AGO 1 h after last injection; §§p < 0.01 AGO 16 h after last injection vs. AGO 1 h after last injection (n =8 rats/group).

Fig 3: Time-dependent fluctuation of CREB, aCaM kinase II, MAPK/Erk1/2 in HPC nuclei. Modulation by chronic agomelatine. Expression levels of CREB (A), aCaM kinase II (C), MAPK/Erk1 (E) and MAPK/Erk2 (G) and phosphorylation levels of CREB at Ser133 (B), aCaM kinase II at Thr286 (D), MAPK/Erk1 at Thr202/204 (F) and MAPK/Erk2 at Thr202/204 (H) in nuclear fraction of HPC from rats chronically treated with vehicle (VEH, black bar) or agomelatine (AGO, white bar) and sacrificed 1 or 16 h after the last injection. Data expressed as above. Statistics as above. **p < 0.01 VEH 16 h after last injection vs. VEH 1 h after last injection; §§§p < 0.001 AGO 16 h after last injection vs. AGO 1 h after last injection (n =8 rats/group).

Fig 4: DJ-1 signalling in vascular smooth muscle cells. DJ-1 modulates VSMCs phenotypic switch DJ-1/ERK/KLF4 signalling pathway

Fig 5: DJ-1 deficiency promotes VSMCs phenotypic switch via increasing KLF4 expression in an ERK-dependent way. A, Immunofluorescence staining of KLF4 expression in aortic root sections of ApoE–/–DJ-1–/– mice and ApoE–/–DJ-1+/+ mice. Dashed line denotes the plaque necrotic core region. L indicates the lumen; A, atheroma. (DAPI = blue staining of nuclei; scale bar = 50 µm) (B,C) Western blot and qRT-PCR analysis of KLF4 protein expression in VSMCs from DJ-1–/– and DJ-1+/+ mice (n = 5; **, P < 0.01 versus DJ-1+/+). D, Western blot analysis protein expression in VSMCs from DJ-1–/– and DJ-1+/+ mice (n = 5). E, Western blot analysis of the expression of contractile (a-SMA and calponin) and synthetic proteins (vimentin, osteopontin and tropoelastin) in DJ-1+/+ VSMCs and DJ-1–/– VSMCs treated with KLF4 siRNA and SCH772984 (n = 5)