Journal: Cellular and molecular life sciences : CMLS

Article Title: Dusp26 phosphatase regulates mitochondrial respiration and oxidative stress and protects neuronal cell death

doi: 10.1007/s00018-022-04162-z

Figure Lengend Snippet: Increased expression and release of HtrA2 and p-HtrA2 from the mitochondria upon loss of Dusp26. A IHC staining of SNpc region showing expression of pS142-HtrA2 (left panel) or HtrA2 (right panel). Quantification of the number of p-HtrA2-positve cells per 1000 cells is presented. n = 5 mice/group. Scale bars p-HtrA2, 10 µm, HtrA2, 20 µm. B IHC staining of SNpc region showing colocalization of HtrA2 with TH in dusp26−/− mice. Quantification of the data is presented. n = 5 mice/group. Scale bars 20 µm. C Immunoblotting analyses of MB cytoplasmic and mitochondrial fractions showing the increased expression of HtrA2 in the cytoplasmic fraction isolated from the mutant mice. The expression of Hsp60 and Hsc70 indicate organelle purity. Quantification of the HtrA2 in the cytoplasmic fraction relative to WT is indicated in the right panel. n = 4–5 mice/group. D Immunoblotting analyses to detect Xiap expression in the MB and cortex of WT and dusp26−/− mice. β-actin is loading control. Quantification of the data relative to WT are presented in the right panel. n = 4–5 mice/group. In all relevant panels, bars are mean ± SEM. p values are indicated in each panel

Article Snippet: The following antibodies were used for IHC staining: Abcam: Dusp26 (b96429); Dako: GFAP (Z0334); Millipore: NeuN (MAB377), TH (AB152); Wako; Iba1 (019–19741); Sigma: TH (T2928); Santa Cruz Biotech: p-p38 (sc-58480), ERK1 (sc-94); Bioss: 8-OHDG (bs1278R); Assay Biotech: HtrA2 (B8515); CUSABIO: pS142-HtrA2 (CSB-PA008883).

Techniques: Expressing, Immunohistochemistry, Western Blot, Isolation, Mutagenesis, Control