Journal: Purinergic Signalling

Article Title: Up-regulation of P2X7 receptors mediating proliferation of Schwann cells after sciatic nerve injury

doi: 10.1007/s11302-015-9445-8

Figure Lengend Snippet: Expression of P2X7R ir in longitudinal sections of normal sciatic nerves. a–c show co-localization of P2X7R ir (red) and S100Aβ ir (green). Note an arrowhead showing a fibre-like structure and an arrow showing a trapezoid structure in a and b; c is the merged image of a and b. Note an arrow indicating a trapezoid structure double labelled by both P2X7R and S100β antibodies (yellow). d–f show co-localization of P2X7R ir (red) and Tuj-1 ir (green). Note an arrow showing a trapezoid structure in d and an arrow showing an axon in e. f is the merged image of d and f; note an arrow indicating a green axon passing through the middle of five trapezoid structures. g–i show co-localization of P2X7R ir (red) and p75NTR ir (green). Note an arrow showing a fibre-like structure in g and h. i is the merged image of g and h. An arrow indicates a double-labelled non-myelinating Schwann cell with P2X7R and p75NTR antibodies (yellow). j–l show co-localization of P2X7R ir (red) and MBP ir (green). Note an arrow showing a fibre-like structure with P2X7R ir, which was not labelled by MBP in l. m–o show co-localization of P2X7R ir (red) and CASPR ir (green). Note that no colocalization of P2X7R ir and CASPR ir was observed. Scale bars in a–c and g–i = 100 μm; scale bars in d–f = 50 μm

Article Snippet: The sections were washed 3–5 min in PBS, and then preincubated in a blocking solution (10 % normal bovine serum, 0.2 % Triton X-100, 0.4 % sodium azide in 0.01 mol/L PBS, pH 7.2) for 30 min followed by incubation with the primary antibodies: P2X7R (1:1000), Alomone, rabbit polyclonal, APR-004; S100β (1:400), Tuj-1 (1:200), p75NTR, PCNA (1:200), MBP (1:200), mouse monoclonal antibodies from Boster, CASPR (1:400) mouse monoclonal antibodies from Santa Cruz, at room temperature overnight.

Techniques: Expressing

Journal: bioRxiv

Article Title: The C-terminal domain of Hsp70 is responsible for paralog-specific regulation of ribonucleotide reductase

doi: 10.1101/2022.02.08.479504

Figure Lengend Snippet: (A-C) Cells expressing either Ssa1, 2, 3 or 4 as their sole Ssa and HA-tagged Rnr1/Rnr2/Rnr4 were grown to exponential phase and were either left untreated or were treated with 200 mM HU for 3 hrs. HA-RNR complexes were immunoprecipitated with anti-HA magnetic beads and were subjected to SDS-PAGE and analyzed by immunoblotting with anti-HA antibodies to detect the RNR subunits or anti-Ydj1 antibodies to detect Ydj1. (D) Interaction between RNR and Ydj1 (Ydj1/RNR) was calculated by quantitating bands from three replicate experiment. Each value represents the mean ± SD (n = 3). Statistical significance between samples was calculated ANOVA. (∗∗∗∗ p < 0.01).

Article Snippet: Proteins were detected using the following antibodies; anti-HA tag (Thermo #26183), Anti-FLAG tag (Sigma, #F1365), anti-PGK1 (Thermo # PA5-28612), anti-Ydj1 (StressMarq #SMC-166D).

Techniques: Expressing, Immunoprecipitation, Magnetic Beads, SDS Page, Western Blot