mouse anti-post synaptic density 95 protein (psd95) 1:20000 1h  (Millipore)

Journal: PLoS ONE

Article Title: Torsin A Localization in the Mouse Cerebellar Synaptic Circuitry

doi: 10.1371/journal.pone.0068063

Figure Lengend Snippet: Western blot of TA, V1, V2, VGAT proteins in cerebellar synaptosomal fractions (SYN) from P14 mice. Synaptosomal preparations are enriched of post synaptic density protein (PSD95) and Synapsin-I (Syn-I) in comparison to equal amount (ACT) of total cerebellar protein extract (LYS). TA is clearly present in synaptosomal enriched preparations. B ) Cerebellar sagittal section immunostained for Cb (green), V1 (cyan) and TA (red). In the ML, anti-V1 specifically labels PF-terminals contacting PC-spines. C – E ) The high magnification of the inset in B is reproduced in different merge images and by the relative split channels. C ) Merge image of Cb and V1 staining plus the colocalization mask Cb/V1 (mCb/V1 in white); it represents the negative CTR (CTR-). D ) Merge image of TA and Cb staining plus the colocalization mask TA/Cb (mTA/Cb in white), highlighting TA expression in PC-spines. E ) Merge image of TA and V1 staining plus the colocalization mask TA/V1 (m TA/V1 in white), indicating the presence of TA also in this synaptic terminal. F ) Quantitative colocalization analysis shows the mean overlap coefficients of TA/Cb (gray column) and TA/V1 (black column) significantly different from the negative CTR Cb/V1 (white column). The small insets (white boxes in C–D-E) are high magnifications of a representative PC-spine (green) contacted by a PF synaptic terminal (cyan) with the appropriate colocalization mask (white). * p < 0.05. Data are represented as mean ± SEM. Scale bars in B, 10 µm; in C–D-E, 5 µm; in F, 1 µm.

Article Snippet: 131011), rabbit anti-Synapsin-I 1:1000 for 1 ON (Novus Biologicals NB300-104), mouse anti-post synaptic density 95 protein (PSD95) 1:20000 1h (Millipore, cod.

Techniques: Western Blot, Staining, Expressing

Journal: Journal of neurochemistry

Article Title: PI3K modulates estrogen-dependent facilitation of colon-to-urethra cross-organ reflex sensitization in ovariectomized female rats.

doi: 10.1111/j.1471-4159.2010.06577.x

Figure Lengend Snippet: Fig. 2 Effects of intrathecal ICI 182780 and LY294002 pre-treatments on estradiol- dependent up-regulation of phosphorylated Akt (pAkt) and phosphorylated NR2B (pNR2B) in left lumbosacral (L6–S2) dorsal horn tissue of ovariectomized rats. Six hours after subcutaneous estradiol injec- tions (OVXE), expression levels of (a) pAkt and (b) pNR2B both increased when com- pared with vehicle injections (OVXV, **p < 0.01 to OVXV, n = 4). Pre-treatments with ICI 182780 (ICI + OVXE) and LY294002 (LY + OVXE), 30 min before estradiol, reversed the increases of pAkt and pNR2B expression caused by estradiol injections (##p < 0.01 to OVXV, n = 4). (c) Co-immunoprecipitation analysis of left lumbosacral (L6–S2) dorsal horn tissue obtained from OVXV and OVXE animals. Immunoblotting (input) in the left column shows increases in expression levels of pAkt, PSD95, and pNR2B in OVXE when compared to OVXV rats. Immunoprecipita- tion blotting (IP) in the right column shows an increment of PSD95 and pNR2B immu- noprecipitation with anti-pAkt antibody in crude membrane extract from the OVXE but not the OVXV group.

Article Snippet: Binding proteins were eluted with SDS–PAGE sample buffer at 95 C. Proteins were separated by SDS–PAGE, transferred to nitrocellulose membranes electrophoretically, and detected using rabbit polyclonal anti-pAkt, mouse monoclonal anti-post-synaptic density protein (PSD95) (Santa Cruz), and rabbit polyclonal antiNR2B (Millipore).

Techniques: Expressing, Immunoprecipitation, Western Blot, Membrane