Journal: PLoS ONE

Article Title: Age-Related Nuclear Translocation of P2X6 Subunit Modifies Splicing Activity Interacting with Splicing Factor 3A1

doi: 10.1371/journal.pone.0123121

Figure Lengend Snippet: ( a-c ) Immunohistochemical analysis of hippocampus sections from 8 month-old mice. Cells were labeled with antibodies against P2X6 ( a ), P2X4 ( b ) and P2X2 ( c ) subunits. Insets depict enlarged views (40X magnification) of CA3 hippocampal region showing a dotted pattern for P2X6 into the nucleus in addition to cytoplasmic staining ( a ), somatodendritic stainining for P2X4 ( b ) and axonal and cytoplasmic staining for P2X2( c ). Scale bar 200 μm. ( d-f ) Immuno-electron microscopy analysis of a CA3 hippocampal neuron from 8 months-old wild type mouse labeled with anti-P2X6 antibody. The staining is observed both in the cytoplasm (cyt, white arrows) and into the nucleus (nuc, white asterisks). Scale bar 2 μm. ( d-e ) Inset of the area indicated in ( e ) showing a 3.5 X magnification of this indicated area. P2X6 positive regions are mainly located in patches added to the inner nuclear membrane ( e ).

Article Snippet: Brain sections were pretreated for 1 h with 1% bovine serum albumin (BSA) (Sigma-Aldrich), 5% FBS, and 0.2% Triton X-100 (Sigma-Aldrich) in PBS, and subsequently incubated with rabbit polyclonal antibodies against P2X6 (Alomone Labs, 1/50), P2X4 (Alomone Labs, 1/500), and/or P2X2 (Alomone Labs, 1/200) receptors.

Techniques: Immunohistochemical staining, Labeling, Staining, Immuno-Electron Microscopy

Journal: PLoS ONE

Article Title: Age-Related Nuclear Translocation of P2X6 Subunit Modifies Splicing Activity Interacting with Splicing Factor 3A1

doi: 10.1371/journal.pone.0123121

Figure Lengend Snippet: ( a ) Western blot analysis for expression of P2X2, P2X4 and P2X6 proteins in mice (from 1 to 18 month-old) ( b ) Graph represent P2X6, P2X2 and P2X4 levels relative to 1 month-old mice. Levels for P2X6 expression is on average 2.58±0.14-fold higher, P2X4 subunit only increases 1.45±0.19-fold at 8 month, and P2X2 level remains invariable (mean±s.e.m, n = 7 mice, * P<0.05, ** P<0.01 One-way ANOVA with Dunnett post-hoc test using 1 month-old mice data as control). ( c,d ) Immunohistochemical analysis of brain section from 1 ( c ) and 8 months-old ( d ) mice. Immunohistochemistry against P2X6 subunits (brown) and detection of senescence-associated β-galactosidase activity (SA-βGal, blue) in hippocampal slices. Scale bar 200 μm. Insets depict enlarged views (40X magnification) of delimited area. ( e ) Quantification of both P2X6 and/or SA-βGal positive cells in hippocampal slices from 1 month-old and 8 month-old mice (mean±s.e.m., n = 3 mice, P = 0.72 unpaired Student’s t-test between same age).

Article Snippet: Brain sections were pretreated for 1 h with 1% bovine serum albumin (BSA) (Sigma-Aldrich), 5% FBS, and 0.2% Triton X-100 (Sigma-Aldrich) in PBS, and subsequently incubated with rabbit polyclonal antibodies against P2X6 (Alomone Labs, 1/50), P2X4 (Alomone Labs, 1/500), and/or P2X2 (Alomone Labs, 1/200) receptors.

Techniques: Western Blot, Expressing, Immunohistochemical staining, Immunohistochemistry, Activity Assay