Fig 2: PPARß/d attenuates cerebral vascular smooth muscle cell phenotypic transformation by suppressing p-Elk-1 and p-p90RSK expression levels. Representative images of western blots showing (A) p-Elk-1 and (C) p-p90RSK protein expression in cerebral vessels at 72 h after SAH (n=6). Expression levels of (B) p-Elk-1 and (D) p-p90RSK were significantly downregulated by pretreatment with Ad-PPARß/d at 72 h after SAH (n=6). *P<0.05 vs. Sham; #P<0.05 vs. SAH + Ad-PPARß/d. PPARß/d, peroxisome proliferator-activated receptor ß/d; p-, phosphorylated; Ad, adenovirus; Elk-1, ETS domain-containing protein Elk-1; p90RSK, p90 ribosomal S6 kinase; SAH, subarachnoid hemorrhage; GFP, green fluorescent protein.

Fig 3: T417+ Elk-1 co-localizes with Ubiquitin+ inclusions in human HD tissue.Striatal inclusions identified by Ubiquitin+ immunoreactivity (yellow arrows in Panel A) also contain T417+ Elk-1 immunoreactivity (yellow arrows in Panel B) (patient HD7; scale bars, 50 µm). Green arrows identify inclusions only containing ubiquitin. Red arrows identify inclusions only containing T417+ Elk-1. Higher magnification images of insets (Panels C–H) (scale bars, 20 µm).

Fig 4: Lactate induces TAM Ap-2a activity and CRC progression. a Nuclear translocation of Ap-2a in TAMs was evaluated. Six-week-old C57BL/6 mice were subcutaneously inoculated with MC-38 cells (1.0 × 106 cells per mouse). TAMs isolated at different time points were stained with anti-Ap-2a antibodies (green). Nuclei were visualized by DAPI staining (blue). The total Ap-2a protein in TAMs at different time points was measured by western blotting assays. The nuclear translocation rates of Ap-2a were calculated (n = 4). b Metabolic factors induced the nuclear translocation of Ap-2a in macrophages. MC-38 conditioned medium was roughly fractionated into a protein fraction (CM-1, >3 kD) and a metabolic fraction (CM-2, <3 kD) by molecular size. Bone marrow-derived macrophages were treated with CM-1, CM-2 or control normal medium for 24 h and then stained with anti-Ap-2a antibodies as described in (a) (n = 3). c Lactate stimulated the nuclear translocation of Ap-2a in macrophages. Bone marrow-derived macrophages were treated with lactate (10 mM) or PBS as a control for 24 h. Then, cells were stained with anti-Ap-2a antibodies as described in (a) (n = 3). d, e The mRNA levels of Elk-1 (d) and Sirpa (e) in macrophages treated with lactate (10 mM) or PBS for 24 h were measured (n = 4). f The phagocytosis rates of macrophages treated with lactate (10 mM) or PBS for 24 h were evaluated (n = 3). g Six-week-old male C57BL/6 mice were subcutaneously engrafted with MC-38 cells (1.0 × 106 cells per mouse) and then subcutaneously treated with lactate (50 µmol in 100 µl of PBS per injection) or PBS on days 1, 3, and 5. Tumor volume was measured dynamically (n = 10). h The survival of the mice described in (g) is shown (n = 10, ***P < 0.005; Gehan–Breslow–Wilcoxon test). The data in (a–g) are reported as the mean ± s.e.m. (***P < 0.005; Student’s t-test)

Fig 5: c-Jun plays an important role in the up-regulation of Talin2 mRNA and miR-190 level. a miR-190 is located in the intron region of the Talin2 gene. b Relative Talin2 mRNA expression was detected in UROtsa and UMUC3 cells. *significant difference (*P < 0.05, **P < 0.01, ***P < 0.001). c The promoter activity of Talin2 was evaluated and compared between UROtsa and UMUC3 cells. *significant difference (*P < 0.05, **P < 0.01, ***P < 0.001). d Potential transcriptional factor-binding sites in the Talin2 promoter region (– 887 to +328) analyzed by using the ALGGEN engine online. e The indicated stable transfectants were subjected to Western Blot to determine p-C-Jun Ser73, C-Jun, C-Fos, Ets-1, Elk-1, and JunB levels. GAPDH was used as a protein loading control. f Relative AP-1 activity was detected in UROtsa and UMUC3 cells. *significant difference (*P < 0.05, **P < 0.01, ***P < 0.001). g c-Jun dominant-negative mutant TAM67 and vector control plasmids were transfected into UMUC3 cells, and c-Jun dominant-negative mutant was identified by western blot. h–j Relative Talin2 promoter activity (h), Talin2 mRNA (i) and miR-190 level (j) detected in TAM67 was compared to that in their vector control cells. *significant difference (*P < 0.05, **P < 0.01, ***P < 0.001). k The proposed schematic for the mechanism underlying miR-190 contributes to Human Bladder Cell Transformation