Journal:

Article Title: Calcium signalling through nucleotide receptor P2X1 in rat portal vein myocytes

doi: 10.1111/j.1469-7793.2001.0339c.xd

Figure Lengend Snippet: A, amplified DNA fragments of P2X receptors (lanes 1–7) from rat brain (a) and rat portal vein myocytes (b) were separated on a 2 % agarose gel and visualized by staining with ethidium bromide. Lane 8, RNA from brain and portal vein myocytes in the absence of reverse transcriptase served as a negative control. Numbers on the left indicate molecular size standards in base pairs (bp). For RNA purification and PCR conditions, see Methods. B, immunostaining of P2X receptor subtypes in portal vein myocytes. Myocytes were stained with anti-P2X1 receptor (a) or anti-P2X4 receptor antibody (b) and vizualization was realized with a donkey anti-rabbit IgG FITC-conjugated antibody. In the absence of primary antibodies or after inactivation of the antibodies by their antigen peptides, only a faint background fluorescence was observed (not shown). Typical confocal sections were performed above the nucleus and therefore appeared spherical. Both P2X1 (a) and P2X4 (b) receptor subtypes were distributed throughout the confocal sections with a marked staining of P2X1 receptor subtype at the cell periphery.

Article Snippet: The rabbit anti-P2X1 and anti-P2X4 receptor antibodies (Alomone Labs, Jerusalem, Israel) were directed against polypeptides corresponding to residues 382–399 and 370–388 of the rat P2X1 and P2X4 receptors, respectively.

Techniques: Amplification, Agarose Gel Electrophoresis, Staining, Negative Control, Purification, Immunostaining, Fluorescence

Journal: Experimental neurology

Article Title: Neuroprotective and Neuro-rehabilitative Effects of Acute Purinergic Receptor P2X4 (P2X4R) Blockade after Ischemic Stroke

doi: 10.1016/j.expneurol.2020.113308

Figure Lengend Snippet: Plot indicates the effect of 5-BDBD at various concentrations (10 nM to 30 μM; log10 [nM] = −4.52 to −8) after P2X4R agonist ATP (1 μM) induced [Ca2+]i response. Calculated IC50 was 3.8 × 10−6 M.

Article Snippet: Primary antibodies used in western blot against P2X4R (Cat. 135341AP), BDNF (Cat. 256991AP), Beta-Actin (Cat. HRP-60008), and secondary antibody Anti-mouse (Cat. SA00002–1) were from Protein Tech, Rosemont, IL.

Techniques:

Journal: Experimental neurology

Article Title: Neuroprotective and Neuro-rehabilitative Effects of Acute Purinergic Receptor P2X4 (P2X4R) Blockade after Ischemic Stroke

doi: 10.1016/j.expneurol.2020.113308

Figure Lengend Snippet: Representative western blots are shown in the top panel. Effects of vehicle or 5-BDBD on pro and mature BDNF at (A) 3 days and (B) 30 days after stroke. C) Effects of vehicle and 5-BDBD treatment on pro/mature BDNF ratio 3 and 30 days after stroke. (D) P2X4R expression at 3 and 30 days after stroke were not different; Data are Mean ± SD (n=4–8 mice/group/time point). *p<0.05 for 5-BDBD vs vehicle by Student’s t-test. β-actin was used as internal control to normalize the data.

Article Snippet: Primary antibodies used in western blot against P2X4R (Cat. 135341AP), BDNF (Cat. 256991AP), Beta-Actin (Cat. HRP-60008), and secondary antibody Anti-mouse (Cat. SA00002–1) were from Protein Tech, Rosemont, IL.

Techniques: Western Blot, Expressing, Control

Journal: Experimental neurology

Article Title: Neuroprotective and Neuro-rehabilitative Effects of Acute Purinergic Receptor P2X4 (P2X4R) Blockade after Ischemic Stroke

doi: 10.1016/j.expneurol.2020.113308

Figure Lengend Snippet: Images (top) and quantification (bottom) of P2X4R expression (green) in lba-1 positive cells (red) from immunostaining of brain sections 3 days after stroke. 20x magnification. DAPI (blue) = 4′,6-diamidino-2-phenylindole (nuclear stain). Scale bar 20 μm. Data are Mean ± SD (n=3 mice/group). *p<0.05, 5-BDBD vs. Vehicle; Student’s t-test.

Article Snippet: Primary antibodies used in western blot against P2X4R (Cat. 135341AP), BDNF (Cat. 256991AP), Beta-Actin (Cat. HRP-60008), and secondary antibody Anti-mouse (Cat. SA00002–1) were from Protein Tech, Rosemont, IL.

Techniques: Expressing, Immunostaining, Staining

Journal: Experimental neurology

Article Title: Neuroprotective and Neuro-rehabilitative Effects of Acute Purinergic Receptor P2X4 (P2X4R) Blockade after Ischemic Stroke

doi: 10.1016/j.expneurol.2020.113308

Figure Lengend Snippet: Flow-sorted microglia and monocytes cells from Fig 5 were fixed (2% paraformaldehyde) and analyzed by an ImageStream cytometer. Using the IDEAS software package, all doublets or aggregated cells were excluded by gating on single cells. A) P2X4R and CD11b+ were focused and selected. Images of single cells (~300 cells/sample, N=3 per group) of the bright field and fluorescent channel (P2X4R-FITC and CD11b-APC-eFluor780) were acquired, and overlays were generated. Magnification=60x. B) Ratio of median fluorescence intensity (MFI) of P2X4R/CD11b was significantly reduced after 5-BDBD treatment in both microglia and monocytes. Further, a separate comparison of microglia vs. monocytes P2X4R MFI suggests reduced expression of P2X4R in microglia irrespective of treatment. Data are Mean ± SD; *p<0.05, 5-BDBD vs. Vehicle; #p<0.05, microglia vs. monocytes; Student’s t-test.

Article Snippet: Primary antibodies used in western blot against P2X4R (Cat. 135341AP), BDNF (Cat. 256991AP), Beta-Actin (Cat. HRP-60008), and secondary antibody Anti-mouse (Cat. SA00002–1) were from Protein Tech, Rosemont, IL.

Techniques: Cytometry, Software, Generated, Fluorescence, Comparison, Expressing

Journal: PLoS ONE

Article Title: Staphylococcus aureus Activates the NLRP3 Inflammasome in Human and Rat Conjunctival Goblet Cells

doi: 10.1371/journal.pone.0074010

Figure Lengend Snippet: All three receptors were identified by western blot. Lanes 1–3 represent separate animals. B is the positive control of rat brain. The β-actin blot is for both P2X4 and P2X7 receptors blots.

Article Snippet: The primary antibodies used for Western blots with the following dilutions: 1∶200 for NLRP3 (R&D Systems, Minneapolis, MN), 1∶50 for ASC, and 1∶1000 for caspase 1, (R&D Systems, Minneapolis, MN) 1∶100 for P2X4 and P2X7, 1∶200 for TLR2 and 1∶1000 for IL-1β (R&D Systems, Minneapolis, MN).

Techniques: Western Blot, Positive Control