PKC zeta Antibody - #AF6405

Fig. 5 Crocetin reduced ROS production by inhibiting NOX2 activity. A Effect of crocetin on MNNG-induced ROS generation (red) in SH-SY5Y cells. Cells were pretreated with crocetin (25, 50, 100 μM) for 1 h and then exposed to MNNG (100 μM) for 15 min. Scale bar: 40 μm. B Group quantification of ROS fluorescence intensity in (A) from three independent experiments. C, D Effect of crocetin on MDA level (C) and GSH level (D) in MNNG-induced early stage of parthanatos. E Confocal images of NADPH oxidase subunits p47phox (green) and gp91phox (red) and DAPI (blue). Cells were pretreated with crocetin (100 μM) for 1 h before MNNG (100 μM) stimulation for 15 min. Scale bar: 10 μm. F Mander’s coefficient between gp91 and p47 staining. G Pearson’s correlation coefficient between gp91 and p47 staining. H Co-immunoprecipitation assay shows MNNG-induced binding between p47phox and gp91phox. Cells were pretreated with crocetin (100 μM) for 1 h before MNNG (100 μM) stimulation for 15 min, and cellular lysates were immunoprecipitated using anti-IgG or anti-gp91phox antibodies and immunoblotted using the indicated antibodies. I Effect of crocetin on MNNG-induced phosphorylation of PKCζ, p47phox, and ERK. J–M Group quantification of gp91 (J), p-PKCζ (K), p-p47 (L), p-ERK (M) in (I) from three independent experiments. Significance was determined by one-way ANOVA. *p 

Fig. 6 Downregulation of polarity-related downstream molecules in incisor odontoblasts on day 13.5. a RT-qPCR quantification analy- sis of intercellular junction-related genes and polarity-related down- stream genes. CA-β-catenin mice displayed downregulated expres- sion levels of N-cadherin, ZO-1, Cdc42, Par3, Par6 and aPKC. b Western blot analysis of intercellular junction-related proteins and polarity-related downstream proteins. The fold activation data analy- sis is shown in (c). CA-β-catenin mice exhibited lower expression levels of N-cadherin, ZO-1, Cdc42, Par3, Par6 and aPKC. *p < 0.05, **p < 0.01, ***p < 0.001, n = 6. (Color figure online)