sc 8066 anti disc1 rabbit wb  (Santa Cruz Biotechnology)

Journal: Brain Sciences

Article Title: Moderate-Intensity Intermittent Training Alters the DNA Methylation Pattern of PDE4D Gene in Hippocampus to Improve the Ability of Spatial Learning and Memory in Aging Rats Reduced by D-Galactose

doi: 10.3390/brainsci13030422

Figure Lengend Snippet: Primers used in the Gene expression analysis.

Article Snippet: The blots were blocked in 5% BSA in Tris-Buffered Saline with Tween (TBST) and incubated with rabbit anti-PDE4A antibody (DF 13317, 1:500, Affinity Biosciences Cincinnati, OH, USA), rabbit anti-PDE4B antibody (72096, 1:1000, CST), rabbit anti-PDE4D antibody (DF6535, 1:800, Affinity Biosciences Cincinnati, OH, USA), rabbit anti-PKA antibody (AF5450, 1:800, Affinity Biosciences Cincinnati, OH, USA), rabbit anti-CREB antibody (9197, 1:1000, CST), rabbit anti-p-CREB antibody (9198, 1:800, CST), rabbit anti-BDNF antibody (DF6387, 1:700, Affinity Biosciences Cincinnati, OH, USA), and rabbit anti-GAPDH antibody (5174, 1:1000, CST).

Techniques: Gene Expression

Journal: Brain Sciences

Article Title: Moderate-Intensity Intermittent Training Alters the DNA Methylation Pattern of PDE4D Gene in Hippocampus to Improve the Ability of Spatial Learning and Memory in Aging Rats Reduced by D-Galactose

doi: 10.3390/brainsci13030422

Figure Lengend Snippet: Effects of MIIT on the expression of PDE4A/B/D mRNA and protein in aging rats: ( a ) The expression of PDE4A/B/D mRNA in hippocampus was detected by Real-Time PCR (mean ± SEM, n = 6). ( b – e ) Western blot analysis of PDE4A/B/D protein expression levels (mean ± SEM, n = 3). Independent-Samples t Test, * p < 0.05 vs. D-Gal group, ** p < 0.01 vs. D-Gal group. Control: young group treated with saline, D-Gal Group: aging group induced by D-galactose, D-Gal+E Group: aging group induced by D-galactose that performed treadmill exercise.

Article Snippet: The blots were blocked in 5% BSA in Tris-Buffered Saline with Tween (TBST) and incubated with rabbit anti-PDE4A antibody (DF 13317, 1:500, Affinity Biosciences Cincinnati, OH, USA), rabbit anti-PDE4B antibody (72096, 1:1000, CST), rabbit anti-PDE4D antibody (DF6535, 1:800, Affinity Biosciences Cincinnati, OH, USA), rabbit anti-PKA antibody (AF5450, 1:800, Affinity Biosciences Cincinnati, OH, USA), rabbit anti-CREB antibody (9197, 1:1000, CST), rabbit anti-p-CREB antibody (9198, 1:800, CST), rabbit anti-BDNF antibody (DF6387, 1:700, Affinity Biosciences Cincinnati, OH, USA), and rabbit anti-GAPDH antibody (5174, 1:1000, CST).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control, Saline

Journal: Communications Biology

Article Title: Phosphodiesterase 4D promotes angiotensin II-induced hypertension in mice via smooth muscle cell contraction

doi: 10.1038/s42003-022-03029-0

Figure Lengend Snippet: a Real-time polymerase chain reaction (RT-PCR) was used to measure mRNA expression of PDE4A, PDE4B, PDE4C, and PDE4D in the aorta tissues of control mice and hypertensive mice (fold change vs. one of controls). b Representative western blot showing PDE4D expression in aorta tissues. c Quantification of PDE4D expression normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein. d Representative immunohistochemical staining of PDE4D. Arrows indicate positive areas. e Quantification of the percentage of PDE4D-positive area. n = 4 in control group, n = 8 in hypertensive group. Data are expressed as mean ± standard error of mean (SEM). Two-tailed Student’s t test was performed to compare differences between two groups. * p < 0.05, *** p < 0.001. L lumen.

Article Snippet: The target protein was probed with numerous antibodies: PDE4A (1:1000, Thermo Fisher, Cat#:PA5-115730), PDE4B (1:1000, Cell Signaling Technology, Cat#: 72096S), PDE4C (1:1000, Thermo Fisher, Cat#: PA5-106624), PDE4D (1:1000, Abcam, Cat#: ab171750), AMP-activated protein kinase (AMPK; 1:1000, Cell Signaling Technology, Cat#: 2532S), Phospho-AMPK (1:1000, Cell Signaling Technology, Cat#: 2535S), myosin phosphatase targeting subunit 1 (MYPT1; 1:1000, Cell Signaling Technology, Cat#: 2634S), Phospho-MYPT1 (1:500, Cell Signaling Technology, Cat#: 5163S), MLC (1:1000, Cell Signaling Technology, Cat#: 8505S), and Phospho-MLC (1:1000, Cell Signaling Technology, Cat#: 3675S), respectively.

Techniques: Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Expressing, Control, Western Blot, Immunohistochemical staining, Staining, Two Tailed Test

Journal: Cellular signalling

Article Title: Phosphodiesterase 4 in inflammatory diseases: Effects of apremilast in psoriatic blood and in dermal myofibroblasts through the PDE4/CD271 complex.

doi: 10.1016/j.cellsig.2016.01.007

Figure Lengend Snippet: Fig. 3. PDE4/CD271 expression in healthy and pathologic dermis. (A) PDE4A, PDE4B, PDE4C, PDE4D isoforms and CD271 in dermis from healthy controls and patients with psoriasis, AD, and DLE were detected by IHC. Fast red was used as chromogen. Bar (shown in top left image = 200 μm) (B) The expression intensity of PDE4A, PDE4B, PDE4C, PDE4D isoforms and CD271 in (A) was quantitatively measured using ImageJ software as described in the Materials and Methods. Relative staining intensities are shown, normalized to intensities in healthy dermis for each individual antibody.

Article Snippet: The blots were blocked for 2 h in blocking buffer (PBS buffer, pH 7.4 with 0.2% Tween 20 and 5% nonfat milk) and incubated with rabbit polyclonal anti-human PDE4A antibody (1:1000; Proteintech Group, Chicago, IL), rabbit polyclonal anti-human PDE4B antibody (1:300; Santa Cruz Biotechnology), rabbit polyclonal anti-human PDE4C antibody (1:300; Santa Cruz Biotechnology), rabbit polyclonal anti-human PDE4D antibody (1:300; Santa Cruz Biotechnology) or mouse monoclonal -SMA (1:3000; Sigma-Aldrich) or mouse monoclonal anti-β-actin (1:1000; AC C EP TE D M AN U SC R IP T Sigma) overnight at 4oC.

Techniques: Expressing, Software, Staining

Journal: Cellular signalling

Article Title: Phosphodiesterase 4 in inflammatory diseases: Effects of apremilast in psoriatic blood and in dermal myofibroblasts through the PDE4/CD271 complex.

doi: 10.1016/j.cellsig.2016.01.007

Figure Lengend Snippet: Fig. 4. PDE4/CD271 expression in fibroblasts and myofibroblasts in vitro. (A) α-SMA expression in DF and DM obtained after treatment with diluent (control) or TGF-β1 (1 ng/mL) for 6 days by immunofluorescence in situ. Cell nuclei were counterstained with DAPI (blue); bar = 20 μm. (B) After diluent (control) or TGF-β1 treatment, levels of α-SMA were determined by Western blot analysis. Vinculin was used as a loading control. (C) Levels of PDE4A, PDE4B, PDE4C, and PDE4D mRNA in DF and DM were determined by RT-PCR analysis. Bar graph shows the average densitometry values normalized to β-actin. (D) PDE4 isoform expression in DF and DM was determined by Western blot analysis. β-Actin was used as a loading control. Bar graphs show the average densitometry values normalized to β-actin (fold-expression). (E) DF and DM protein extracts were immunoprecipitated with CD271 antibody and immunoblotted with PDE4A, PDE4B, PDE4C, and PDE4D antibodies and with PAS-only as control.

Article Snippet: The blots were blocked for 2 h in blocking buffer (PBS buffer, pH 7.4 with 0.2% Tween 20 and 5% nonfat milk) and incubated with rabbit polyclonal anti-human PDE4A antibody (1:1000; Proteintech Group, Chicago, IL), rabbit polyclonal anti-human PDE4B antibody (1:300; Santa Cruz Biotechnology), rabbit polyclonal anti-human PDE4C antibody (1:300; Santa Cruz Biotechnology), rabbit polyclonal anti-human PDE4D antibody (1:300; Santa Cruz Biotechnology) or mouse monoclonal -SMA (1:3000; Sigma-Aldrich) or mouse monoclonal anti-β-actin (1:1000; AC C EP TE D M AN U SC R IP T Sigma) overnight at 4oC.

Techniques: Expressing, In Vitro, Control, In Situ, Western Blot, Reverse Transcription Polymerase Chain Reaction, Immunoprecipitation