Journal: Neural Regeneration Research

Article Title: Prolonged intermittent theta burst stimulation restores the balance between A2AR- and A1R-mediated adenosine signaling in the 6-hydroxidopamine model of Parkinson’s disease

doi: 10.4103/nrr.nrr-d-23-01542

Figure Lengend Snippet: Figure 4 ｜Effects of prolonged iTBS on expression of crude membrane (P2) receptors in the caudoputamen. (A) Representative support membranes showing density of 70-kDa (P2X7R) and 37-kDa (GAPDH) bands and histograms of immunoblot analysis showing P2X7R protein abundance. (B) Representative support membranes showing density of 70-kDa (P2Y1R) and 37-kDa (GAPDH) bands and histograms of immunoblot analysis showing P2Y1R protein abundance. (C) Representative support membranes showing density of 70-kDa (P2Y12R) and 37-kDa (GAPDH) bands and histograms of immunoblot analysis showing P2Y12R protein abundance stimulation. (D) Representative support membranes showing the density of 70-kDa (P2Y13R) and 37-kDa (GAPDH) bands, as well as histograms of immunoblot analysis displaying P2Y13R protein abundance in crude synaptosomal fractions of the CPu of sham-stimulated and iTBS-stimulated animals, 1 and 3 weeks after stimulation. (E) ADP in crude synaptosomal fraction from CPu from Sham- and iTBS-stimulated animals 1 and 3 weeks after stimulation. Bars show mean activity (nmol Pi/mg/min) ± SEM from n = 5 determinations performed in duplicate. The number at the bottom of the graphs represents the number of individual animals included in analysis. *P < 0.05, **P < 0.01 (unpaired Student’s t-test). All experiments were repeated at least twice. CPu: Caudoputamen; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; iTBS: intermittent theta burst stimulation; ns: not significant.

Article Snippet: After Table 1 | Primary and secondary antibodies used in this study Antibody Host organism and clonality Dilution Supplier Cat# RRID TH Rabbit, polyclonal 1:2000WB, 1:500IHC Millipore, Darmstadt, Germany AB152 AB_390204 GFAP Rabbit, polyclonal 1:500IHC, 1:7000WB Santa Clara, CA, USA Z0334 AB_10013382 Iba-1 Goat, polyclonal 1:400IHC, 1:500WB Abcam, Cambridge, MA, USA AB5076 AB_2224402 CD73, rNu-9L(I4,I5) Rabbit, polyclonal 1:200IHC Ectonucleotidases-ab.com, IL-1β Rabbit, polyclonal 1:500DB Fine Test, Wuhan China FNab04209 TNF-α Rabbit, polyclonal 1:500DB Thermo Fisher Scientific, Waltham, MA, USA PA1-40281 AB_2204371 P2X7R Rabbit, polyclonal 1:1000WB Alomone Labs, Jerusalem, Israel AAR-004 AB_2040068 P2Y1R Rabbit, polyclonal 1:1000WB Alomone Labs, Jerusalem, Israel AAR-021 AB_10919250 P2Y12R Rabbit, polyclonal 1:1000WB Alomone Labs, Jerusalem, Israel AAR-012 AB_2040074 P2Y13R Rabbit, polyclonal 1:1000WB Alomone Labs, Jerusalem, Israel AAR-017 AB_2040076 CD73 Rabbit, polyclonal 1:1000WB Cell Signaling Technology, Danvers, MA, USA 13160 AB_2716625 ADA Rabbit, polyclonal 1:1000WB Thermo Fisher Scientific PA5-51572 AB_2637694 A1R Rabbit, polyclonal 1:1000WB Alomone Labs AAR-009 A2AR Rabbit, polyclonal 1:1000WB Thermo Fisher Scientific PA1-042 AB_2257858 p-AMPK Rabbit, polyclonal 1:2000WB Cell Signaling Technology 2535 AB_331250 t-AMPK Rabbit, polyclonal 1:2000WB Cell Signaling Technology 2532 AB_330331 D1DR Rabbit, polyclonal 1:500WB Abcam AB81296 AB_2814742 D2DR Rabbit, polyclonal 1:500WB Fine Test FNab02533 GAPDH Rabbit, polyclonal 1:2000WB Thermo Fisher Scientific PA1-987 AB_2107311 Goat anti-rabbit IgG, HRP-conjugated Goat, polyclonal 1:30000WB Abcam AB6721 AB_955447 Rabbit anti-goat IgG, HRP-conjugated Rabbit, polyclonal 1:10000WB R and D Systems , Minneapolis, MN, USA HAF017 AB_562588 DB: Dot blot; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GFAP: glial fibrillary acidic protein; HRP: horseradish peroxidase; Iba-1: ionized calcium binding adaptor molecule 1; IL-1β: interleukin-1β; TH: tyrosine hydroxylase; TNF-α: tumor necrosis factor-α; p-AMPK: phospho-adenosine monophosphate kinase, t-AMPK: total-adenosine monophosphate kinase; D1DR: dopamine 1 receptor, D2DR: dopamine 2 receptor; WB: western blotting; IHC: immunohistochemistry.

Techniques: Expressing, Membrane, Western Blot, Quantitative Proteomics, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Role of α2-Adrenoceptor Subtypes in Suppression of L-Type Ca 2+ Current in Mouse Cardiac Myocytes

doi: 10.3390/ijms22084135

Figure Lengend Snippet: BRL 44408, a specific antagonist of the α2A isoform, did not affect the guanabenz-induced suppression of I CaL . ( a , b ) Representative I CaL recordings and time-course of peak I CaL density values in the presence of guanabenz (guan) and BRL 44408. Horizontal bars denote the protocol of the drug applications. ( c ) BRL 44408 did not induce a significant rightward shift of dose-dependent inhibition of I CaL by guanabenz. Curves represent the Hill’s fits with parameters indicated in the text.

Article Snippet: Proteins were separated in 10% SDS-PAGE, transferred to nitrocellulose membranes (sc-3724, 0.45 μm, Santa Cruz Biotechnology) and probed with antibodies against α2A (AAR-020, Alomone labs, lot AAR020AN0202), α2B (AAR-021, Alomone labs, lot AAR021AN0202) and α2C (ab46536, Abcam) diluted to 1:100.

Techniques: Inhibition

Journal: International Journal of Molecular Sciences

Article Title: Role of α2-Adrenoceptor Subtypes in Suppression of L-Type Ca 2+ Current in Mouse Cardiac Myocytes

doi: 10.3390/ijms22084135

Figure Lengend Snippet: The mRNA and protein expression levels of the α2-AR isoforms in the mouse left ventricle. ( a ) mRNA expression of the α2 adrenoceptor genes obtained using the RT-qPCR assay. In contrast to adra2B and adrs2C genes the expression of adra2A was not detected (ND). ( b ) Western blots confirmed the expression of α2B and α2C but not α2A receptor proteins in the mouse left ventricular tissue.

Article Snippet: Proteins were separated in 10% SDS-PAGE, transferred to nitrocellulose membranes (sc-3724, 0.45 μm, Santa Cruz Biotechnology) and probed with antibodies against α2A (AAR-020, Alomone labs, lot AAR020AN0202), α2B (AAR-021, Alomone labs, lot AAR021AN0202) and α2C (ab46536, Abcam) diluted to 1:100.

Techniques: Expressing, Quantitative RT-PCR, Western Blot