anti α 2a ar  (Proteintech)

Journal: Nature Communications

Article Title: Specific pharmacological and G i/o protein responses of some native GPCRs in neurons

doi: 10.1038/s41467-024-46177-z

Figure Lengend Snippet: a Effect of the indicated α 2 AR agonists on the net BRET of the G i1 and G oA sensors in CGNs co-transfected with Gα i1 Nluc and Venus Gγ 2 or Gα oA Nluc and Venus Gγ 2 (amounts of cDNA as in Fig. ). b Effect of the indicated α 2 AR agonists on the net BRET of the G i1 and G oA sensors in HEK293 cells co-transfected with the mouse α 2A AR (10 ng, 20 ng or 50 ng), and Gβ 1, Venus Gγ 2 and Gα i1 Nluc or Gα oA Nluc as in Fig. . Saturating concentrations of brimonidine (10 μM), oxymetazoline (100 μM), xylazine (50 μM), clonidine (100 μM), tizanidine (100 μM) were used in ( a , b ). c Detection of α 2 AR in cell membranes of CGNs, mock-transfected HEK293 cells and HEK293 cells transfected with indicated amount of α 2 AR, by western blotting. Values are mean ± SEM normalized as fold of α 2 AR expression in CGNs from four independent experiments. d The pEC 50 of brimonidine in CGNs ( n = 3) and transfected HEK293 cells ( n = 4) with indicated amount of α 2 AR measured by G i1 or G oA sensor. e Percentage of change in BRET ratio between Gα Nluc and Venus Gγ 2 induced by brimonidine between Gα Nluc and Venus Gγ 2 in HEK293 cells ( n = 3) (α 2 AR: 15 ng/well per 96-well plate) or CGNs ( n = 4) for the indicated Gα i1 , Gα i2 , Gα i3 , Gα oA , Gα oB or Gα z sensors (amounts of cDNA as in Fig. ). Values are mean ± SEM from at least three biologically independent experiments each performed in triplicate or quadruplicate in ( a , b , d , e ). a , n = 4; b , n = 3; ( d , e ), CGNs, n = 3; HEK293 cells, n = 4. Data are analysed using one-way ANOVA with a Dunnett’s post-hoc multiple comparison test to determine significance (compared with brimonidine in a , b ). Data are analysed using unpaired t -test (two-tailed) in ( e ). **** p < 0.0001, *** p < 0.001, ** p < 0.01 and not significant (ns). The raw data and p -values are available in source data provided as a Source Data file.

Article Snippet: The blots were incubated with the primary monoclonal antibodies anti-GB1 mAb (1:1000, ab55051, Abcam, Shanghai, China), anti-Gβ 2 rabbit (1:1000, A9643, ABclonal Technology, Wuhan, China), anti-β-actin (1:3000, KM9001T, Sungene Biotech., Tianjin, China), the rabbit polyclonal antibodies anti-CB1 (1:1000, A1447, ABclonal, Wuhan, China), anti-α 2A AR (1:1000, A2809, ABclonal, Wuhan, China), anti-Gβ 1 (1:1000, A1867, ABclonal, Wuhan, China), anti-Gβ 3 (1:1000, A1387, ABclonal, Wuhan, China) and anti-Gβ 5 (1:1000, A4447, ABclonal, Wuhan, China).

Techniques: Transfection, Western Blot, Expressing, Comparison, Two Tailed Test

Journal: Journal of Cellular and Molecular Medicine

Article Title: The α2AR/Caveolin‐1/p38MAPK/NF‐κB axis explains dexmedetomidine protection against lung injury following intestinal ischaemia‐reperfusion

doi: 10.1111/jcmm.16614

Figure Lengend Snippet: Dex reduces lung injury following intestinal I/R via Cav‐1‐dependent p38MAPK/NF‐κB pathway inactivation. A, Comparison of the down‐regulation for Cav‐1 protein using three anti‐Cav‐1 shRNA constructs. * P < .05 compared with the mock group. Pre‐injections were applied four days before intestinal I/R injury. B, RT‐qPCR of Cav‐1 mRNA expression and Western blot analysis of Cav‐1, p38, p‐p38, p‐p65 and p65 expression in rat lung tissues with Cav‐1 overexpression or knockdown before intestinal I/R injury. C, ELISA detection of TNF‐α expression in rat lung tissues with Cav‐1 overexpression or knockdown before intestinal I/R injury. D, ELISA detection of IL‐1β expression in rat lung tissues with Cav‐1 overexpression or knockdown before intestinal I/R injury. E, ELISA detection of IL‐6 expression in rat lung tissues with Cav‐1 overexpression or knockdown before intestinal I/R injury. F, Lung injury score of rat lung tissues with Cav‐1 overexpression or knockdown before intestinal I/R injury. G, Apoptosis rate of rat lung tissues with Cav‐1 overexpression or knockdown before intestinal I/R injury. * P < .05 vs sham group, # P < .05 vs I/R + oe‐NC group. H, Western blot analysis of Cav‐1, p38, p‐p38, p‐p65 and p65 in rat lung tissues upon Dex treatment and/or Cav‐1 knockdown prior to intestinal I/R injury. I, ELISA detection of TNF‐α expression in rat lung tissues upon Dex treatment and/or Cav‐1 knockdown prior to intestinal I/R injury. J, ELISA detection of IL‐1βexpression in rat lung tissues upon Dex treatment and/or Cav‐1 knockdown prior to intestinal I/R injury. K, ELISA detection of IL‐6 expression in rat lung tissues upon Dex treatment and/or Cav‐1 knockdown prior to intestinal I/R injury. L, Lung injury score of rat lung tissues upon Dex treatment and/or Cav‐1 knockdown prior to intestinal I/R injury. M, Apoptosis rate of rat lung tissues upon Dex treatment and/or Cav‐1 knockdown prior to intestinal I/R injury, detected by TUNEL staining. * P < .05 vs sham group, # P < .05 vs I/R + Dex +sh‐NC group. Data among multiple groups were analysed by one‐way ANOVA with Tukey's post hoc test

Article Snippet: The PVDF membrane was blocked in 5% skimmed milk at room temperature for 1 hours and incubated overnight at 4°C with the diluted primary antibodies: rabbit anti‐α 2A ‐AR (1:1000, 14266‐1‐AP, Proteintech ProteinTech Group), rabbit anti‐Cav‐1 (1:1000, ab32577, Abcam Inc.), rabbit anti‐p38 (1:1000, ab31828, Abcam), rabbit anti‐phosphorylated (p)‐p38 (1:1000, ab4822, Abcam), rabbit anti‐NF‐κBp65 (1:1000, ab16502, Abcam), rabbit anti‐NF‐κB‐p‐p65 (1:1000, ab86299, Abcam) and rabbit anti‐Actin (1:1000, ab179467, Abcam).

Techniques: Comparison, shRNA, Construct, Quantitative RT-PCR, Expressing, Western Blot, Over Expression, Knockdown, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Staining

Journal: Frontiers in Neuroscience

Article Title: Effects of α2A Adrenoceptors on Norepinephrine Secretion from the Locus Coeruleus during Chronic Stress-Induced Depression

doi: 10.3389/fnins.2017.00243

Figure Lengend Snippet: Intraperitoneal injection of BRL-44408 maleate can reverse the increase of α 2A -AR induced by CUMS in the hypothalamus of the CUMS rats . (A.B ) The protein expression levels of α 2A -AR was significantly increased in the hypothalamus of the CUMS rats, peripheral administration of BRL-44408 maleate significantly decreased the expression levels of α 2A -AR in the CUMS rats. * p < 0.05 vs. the control rats. # p < 0.05 vs. before BRL-44408 maleate in the CUMS group. NS represents no significance.

Article Snippet: Then, the protein components were transferred to polyvinylidene difluoride (PVDF) membranes, and then blocked with 5% BSA in TBST (TBS+0.1% Tween-20) for 1 h, and then immunoblotted overnight at 4°C with primary antibody for α 2A -AR (#14266-1-AP, Proteintech, USA).

Techniques: Injection, Expressing, Control