Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

Article Title: Transfer, analysis and reversion of the fibrous dysplasia cellular phenotype in human skeletal progenitors

doi: 10.1359/jbmr.091036

Figure Lengend Snippet: Table III

Article Snippet: PDE4A , hCG28500 , Hs00183479_m1.

Techniques:

Journal: Frontiers in Aging

Article Title: Microvascular Sex- and Age- Dependent Phosphodiesterase Expression

doi: 10.3389/fragi.2021.719698

Figure Lengend Snippet: PDE4 protein expression in skeletal muscles arterioles and venules. (A) The representative immunoblotting using non-selective PDE4 antibody shows PDE4 protein expressed in arterioles and venules isolated from rat abdominal skeletal muscles. Rat brain was used as a positive control. The expression of β-actin protein was used as a loading control for corresponding vessels. Art: arteriole; Ven: venule. (B) The representative immunofluorescence imaging was acquired using scanning confocal microscopy. The sectioned rat abdominal skeletal muscle was stained with PDE4A primary and Alexa-488 (green color) secondary antibodies (i) as well as CD31 (an endothelial marker) primary and Alexa-568 (red color) secondary antibody (ii) . The images of (i) and (ii) were superimposed (iii) , in which the yellow color indicates co-localization of PDE4A and CD31. The section in the small box of image c was enlarged (iv) , showing the co-localization of PDE4A and CD31 molecules. The transmitted light image (v) shows the venule of skeletal muscle tissue. The scale bar represents 50 µm for all the images except the image (iv) .

Article Snippet: TaqMan® Gene Expression Assay (Applied Biosystems™, CA) was used for PDE real time PCR analysis: PDE1A, Rn00578422_m1; PDE1B, Rn00575591_m1; PDE1C, Rn00579334_m1; PDE2A, Rn00579346_m1; PDE3A, Rn00569192_m1; PDE3B, Rn00568191_m1; PDE4A, Rn00565354_m1; PDE4B, Rn00566785_A1; PDE4D, Rn00566798_m1; and PDE5A, Rn00592185_m1. ß-actin (Rn00667869_m1) served as the internal control because it has been accepted that ß-actin mRNA expression remains relatively constant under a variety of conditions.

Techniques: Expressing, Western Blot, Isolation, Positive Control, Immunofluorescence, Imaging, Confocal Microscopy, Staining, Marker

Journal: Scientific Reports

Article Title: Phosphodiesterase type 4 inhibition enhances nitric oxide- and hydrogen sulfide-mediated bladder neck inhibitory neurotransmission

doi: 10.1038/s41598-018-22934-1

Figure Lengend Snippet: Expression of PDE4A protein within nerve fibers distributed among pig bladder neck smooth muscle bundles. Double-labeling immunofluorescence assay in the pig bladder neck ( A – D ). Bladder neck overall innervation was visualized using the general nerve marker PGP 9.5 (red areas) ( A ). PDE4A immunofluorescence from pig bladder neck reveals immunopositive nerve trunks (green areas), running parallel to the smooth muscle bundles. Same fields ( A , B , E and F ). Immunofluorescence double-labeling for PGP 9.5 and PDE4A in the smooth muscle, demonstrate neuronal co-localization (yellow areas) ( C ). Cell nuclei were counterstained with DAPI (blue areas) ( D ). Scale bars indicate 25 µm. Negative controls showing the lack of immunoreactivity in sections incubated in the absence of the primary antibody ( E and F ) (n = 5). Comparison of the fluorescence of PGP 9.5 and PDE4A, using ImageJ, which shows a major co-localization between PDE4A and PGP 9.5 ( G ). Western blot of smooth muscle membranes from pig bladder neck incubated with PDE4A antibody showing a 118 kDa major band, which corresponded to the expected molecular weight ( H ) (n = 5).

Article Snippet: The tissue was sliced into transversal sections 5 μm thickness and preincubated in 10% normal donkey serum in PB containing 0.3% Triton-X-100, for 2–3 h. Followed the sections were incubated with either goat anti-PDE4 (anti-NPP4 C-15, Santa Cruz Biotechnology Inc. Heidelberg, Germany) at a 1:50 dilution plus a mouse anti-protein gene product 9.5 (anti-PGP 9.5), diluted 1:100 and mouse anti PDE4A (anti-PDE4A H-7, Santa Cruz Biotechnology Inc. Heidelberg, Germany) at a 1:50 dilution plus a rabbit anti-protein gene product 9.5 (anti-PGP 9.5) diluted 1:50 during 48 h at 4 °C.

Techniques: Expressing, Labeling, Immunofluorescence, Marker, Incubation, Comparison, Fluorescence, Western Blot, Molecular Weight

Journal: Scientific Reports

Article Title: Phosphodiesterase type 4 inhibition enhances nitric oxide- and hydrogen sulfide-mediated bladder neck inhibitory neurotransmission

doi: 10.1038/s41598-018-22934-1

Figure Lengend Snippet: Expression of PDE4A protein within nerve fibers distributed among human bladder neck smooth muscle bundles. Double-labeling immunofluorescence assay in the human bladder neck ( A – D ). Bladder neck overall innervation was visualized using the general A marker PGP 9.5 (red areas) ( A ). PDE4A immunofluorescence from human bladder neck reveals immunopositive nerve trunks (green areas), running parallel to the smooth muscle bundles. Same fields ( A , B , E and F ). Immunofluorescence double-labeling for PGP 9.5 and PDE4A in the smooth muscle, demonstrate neuronal co-localization (yellow areas) ( C ). Cell nuclei were counterstained with DAPI (blue areas) ( D ). Scale bars indicate 25 µm. Negative controls showing the lack of immunoreactivity in sections incubated in the absence of the primary antibody ( E and F ) (n = 4). Comparison of the fluorescence of PGP 9.5 and PDE4A, using ImageJ, which shows a major co-localization between PDE4A and PGP 9.5 ( G ). Western blot of smooth muscle membranes from human bladder neck incubated with PDE4A antibody showing a 118 kDa major band, which corresponded to the expected molecular weight ( H ) (n = 4).

Article Snippet: The tissue was sliced into transversal sections 5 μm thickness and preincubated in 10% normal donkey serum in PB containing 0.3% Triton-X-100, for 2–3 h. Followed the sections were incubated with either goat anti-PDE4 (anti-NPP4 C-15, Santa Cruz Biotechnology Inc. Heidelberg, Germany) at a 1:50 dilution plus a mouse anti-protein gene product 9.5 (anti-PGP 9.5), diluted 1:100 and mouse anti PDE4A (anti-PDE4A H-7, Santa Cruz Biotechnology Inc. Heidelberg, Germany) at a 1:50 dilution plus a rabbit anti-protein gene product 9.5 (anti-PGP 9.5) diluted 1:50 during 48 h at 4 °C.

Techniques: Expressing, Labeling, Immunofluorescence, Marker, Incubation, Comparison, Fluorescence, Western Blot, Molecular Weight

Journal: Journal of Cancer

Article Title: PDE4a predicts poor prognosis and promotes metastasis by inducing epithelial-mesenchymal transition in hepatocellular carcinoma

doi: 10.7150/jca.24079

Figure Lengend Snippet: Demographic information and clinical feature of 210 HCC patients

Article Snippet: The siRNA sequence against PDE4a (Catalog No.: sc-41596) and relative srambled siRNA sequence (Catalog No.: sc-37007) were from Santa Cruz Biotechnology (CA, USA), which were transfected into MHCC97h cells by siRNA transfection reagent (Catalog No.:sc-29528, Santa Cruz Biotechnology, USA) respectively to be MHCC97h PDE4a siRNA cells and MHCC97h Scr siRNA cells.

Techniques: Infection

Journal: Journal of Cancer

Article Title: PDE4a predicts poor prognosis and promotes metastasis by inducing epithelial-mesenchymal transition in hepatocellular carcinoma

doi: 10.7150/jca.24079

Figure Lengend Snippet: The PDE4a was up-regulated in HCC tissues and HCC cell lines. A . The representative picture of PDE4a IHC staining in adjacent liver tissues (a) and HCC tissues (b). There was significantly more PDE4a expression in HCC tissues in contrast to adjacent liver tissues; B . Mann-Whitney U test demonstrated that PDE4a expression in HCC tissues was apparently higher than one in adjacent liver tissues; C . Both qRT-PCR and Western immunoblotting showed that there was the higher level of PDE4a expression in 5 kinds of HCC cell line (Huh7, Hep3B, MHCC97h, SK-Hep1 and PLC/PRF/5) than human liver cell line LO2. Among 5 kinds of HCC cell lines, MHCC97h cells had the highest level of PDE4a expression, while there was the least PDE4a expression in Huh7 cells.

Article Snippet: The siRNA sequence against PDE4a (Catalog No.: sc-41596) and relative srambled siRNA sequence (Catalog No.: sc-37007) were from Santa Cruz Biotechnology (CA, USA), which were transfected into MHCC97h cells by siRNA transfection reagent (Catalog No.:sc-29528, Santa Cruz Biotechnology, USA) respectively to be MHCC97h PDE4a siRNA cells and MHCC97h Scr siRNA cells.

Techniques: Immunohistochemistry, Expressing, MANN-WHITNEY, Quantitative RT-PCR, Western Blot

Journal: Journal of Cancer

Article Title: PDE4a predicts poor prognosis and promotes metastasis by inducing epithelial-mesenchymal transition in hepatocellular carcinoma

doi: 10.7150/jca.24079

Figure Lengend Snippet: Univariate and multivariate analyses of predictive factors in HCC patients with liver resection

Article Snippet: The siRNA sequence against PDE4a (Catalog No.: sc-41596) and relative srambled siRNA sequence (Catalog No.: sc-37007) were from Santa Cruz Biotechnology (CA, USA), which were transfected into MHCC97h cells by siRNA transfection reagent (Catalog No.:sc-29528, Santa Cruz Biotechnology, USA) respectively to be MHCC97h PDE4a siRNA cells and MHCC97h Scr siRNA cells.

Techniques: Infection

Journal: Journal of Cancer

Article Title: PDE4a predicts poor prognosis and promotes metastasis by inducing epithelial-mesenchymal transition in hepatocellular carcinoma

doi: 10.7150/jca.24079

Figure Lengend Snippet: Enforced expression of PDE4a induced EMT phenotype of Huh7 cell. A . Transfection of PDE4a expressing plasmid resulted in up-regulation of PDE4a in Huh7 cells; B . As assessed by MTT assay, enhanced expression of PDE4a increased the cell viability of Huh7 cells; C . ELISA assay displayed that there was magnificently more BrdU incorporation found in Huh7 PDE4a cells than Huh7 Vector cells; D . As examined by scratch wound healing assay, cell migration of Huh7 cells was strengthened by over-expression of PDE4a significantly; E . Transwell chamber coated with Matrigel matrix was used to measure the cell invasion ability and it was found that over-expression of PDE4a enhanced invasion capacity of Huh7 cells clearly; F . Enforced expression of PDE4a repressed E-cadherin expression and up-regulated the expression of N-cadherin, TWIST, Vimentin and TWIST in Huh7 cells. Supplementary fig. presented the expression of E-cadherin, N-cadherin, Vimentin and TWIST examined by Western immunoblotting semiquantitatively.

Article Snippet: The siRNA sequence against PDE4a (Catalog No.: sc-41596) and relative srambled siRNA sequence (Catalog No.: sc-37007) were from Santa Cruz Biotechnology (CA, USA), which were transfected into MHCC97h cells by siRNA transfection reagent (Catalog No.:sc-29528, Santa Cruz Biotechnology, USA) respectively to be MHCC97h PDE4a siRNA cells and MHCC97h Scr siRNA cells.

Techniques: Expressing, Transfection, Plasmid Preparation, MTT Assay, Enzyme-linked Immunosorbent Assay, BrdU Incorporation Assay, Wound Healing Assay, Migration, Over Expression, Western Blot

Journal: Journal of Cancer

Article Title: PDE4a predicts poor prognosis and promotes metastasis by inducing epithelial-mesenchymal transition in hepatocellular carcinoma

doi: 10.7150/jca.24079

Figure Lengend Snippet: Knockdown of PDE4a attentuated the malignant behaviors and reversed EMT phenotype in MHCC97h cells. A . As examined by both qRT-PCR and Western immunoblotting assays, PDE4a expression was found significantly decreased by transfection of PDE4a siRNA sequences; B . MTT assay showed that silencing PDE4a leaded to repression of cell viability in MHCC97h cells; C . BrdU ELISA assay displayed that knockdown of PDE4a restrained cell proliferation in MHCC97h cells significantly; D. Invasion capacity of MHCC97h cells was repressed clearly by knockdown of PDE4a; E . Silencing PDE4a was found to inhibit expression of both N-cadherin, Vimentin and TWIST, and increase E-cadherin expression by Western immunoblotting assay. Supplementary fig. displayed the expression of E-cadherin, N-cadherin, Vimentin and TWIST examined by Western immunoblotting semiquantitatively.

Article Snippet: The siRNA sequence against PDE4a (Catalog No.: sc-41596) and relative srambled siRNA sequence (Catalog No.: sc-37007) were from Santa Cruz Biotechnology (CA, USA), which were transfected into MHCC97h cells by siRNA transfection reagent (Catalog No.:sc-29528, Santa Cruz Biotechnology, USA) respectively to be MHCC97h PDE4a siRNA cells and MHCC97h Scr siRNA cells.

Techniques: Knockdown, Quantitative RT-PCR, Western Blot, Expressing, Transfection, MTT Assay, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Genomics

Article Title: Comprehensive Analysis of Transcriptome Sequencing Data in the Lung Tissues of COPD Subjects

doi: 10.1155/2015/206937

Figure Lengend Snippet: Primers for mRNA expression profiling.

Article Snippet: PDE4A , Hs01102342_mH , ACCGCATCCAGGTCCTCCGGAACAT.

Techniques: Expressing, Sequencing