Journal: PLoS ONE

Article Title: 2’,3’-Cyclic Nucleotide 3’-Phosphodiesterases Inhibit Hepatitis B Virus Replication

doi: 10.1371/journal.pone.0080769

Figure Lengend Snippet: (A) The expression of the CNP proteins was studied by immunofluorescence microscopy using the anti-CNP polyclonal rabbit antibody and Alexa Fluor 488-anti-rabbit IgG. The nuclei were counterstained with 4,6-diamidino-2-phenylindole (DAPI). (B) Huh7 and HepG2 cells were cultured without or with 300, 1000 IU/ml of IFNα-2b for 2 days. Cells were harvested and the cell lysates were determined by Western blot analysis. Actin served as a loading control (protein panel). (C) For CNP mRNA quantitative analysis, cDNA pools were synthesized from the 1 μg of total RNA of cells and diluted cDNA was amplified by quantitative real-time PCR. Actin was served as reverse transcription and PCR control. The mRNA% presents the ratio of values of total CNP (tCNP) to that of naive HepG2 cells (n=3).

Article Snippet: Membranes were blocked with 0.05% TBST (Tris-Buffered Saline Tween-20) containing 5% nonfat dry milk and probed with antibodies against CNP (Proteintech 1:3000), β-actin (Abcam, 1:5000), secondary antibodies conjugated to horseradish peroxidase were used and followed by ECL (Merck Millipore) detection.

Techniques: Expressing, Immunofluorescence, Microscopy, Cell Culture, Western Blot, Control, Synthesized, Amplification, Real-time Polymerase Chain Reaction, Reverse Transcription

Journal: PLoS ONE

Article Title: 2’,3’-Cyclic Nucleotide 3’-Phosphodiesterases Inhibit Hepatitis B Virus Replication

doi: 10.1371/journal.pone.0080769

Figure Lengend Snippet: (A) Huh7 cells were transfected with pCDNA5-CNP1 or pCDNA5-CNP2 and harvested at day 2 post transfection. The expression of CNP and tubulin was revealed by immunofluorescence microscopy using the anti-CNP polyclonal rabbit antibody and anti-tublin monoclonal mice antibody, the secondary antibodies were Alexa Fluor 594-conjugated goat anti-rabbit IgG Alexa Fluor 488-conjugated goat anti-mouse or rabbit IgG. The nuclei were counterstained with DAPI. Mitochondria were stained using Mitocapture Tm kit and indicated by red color. White arrow indicates a CNP1-transfected Huh7 cell undergone a morphological change. (B) Huh7 ells in 12-well plate were transfected with 2 μg of plasmids pCDNA5-CNP1 or pCDNA5-CNP2. Cell lysates were determined by Western blot analysis. Actin served as a loading control.

Article Snippet: Membranes were blocked with 0.05% TBST (Tris-Buffered Saline Tween-20) containing 5% nonfat dry milk and probed with antibodies against CNP (Proteintech 1:3000), β-actin (Abcam, 1:5000), secondary antibodies conjugated to horseradish peroxidase were used and followed by ECL (Merck Millipore) detection.

Techniques: Transfection, Expressing, Immunofluorescence, Microscopy, Staining, Western Blot, Control

Journal: PLoS ONE

Article Title: 2’,3’-Cyclic Nucleotide 3’-Phosphodiesterases Inhibit Hepatitis B Virus Replication

doi: 10.1371/journal.pone.0080769

Figure Lengend Snippet: Panels show micrographs of liver biopsy specimens from three HBV-infected patients. Specimens were costained with antibodies against HBV X protein (red) and CNP (green); nuclei were stained with DAPI (blue). These images were collected at 100 magnification and show the representative images. Control micrograph was collected from the non-infectious area. The dashed rectangle in P2 (Patient 2) indicated the HBx positive but CNP negative cells; the real line indicated the both positive area. The enlarge panel exhibits the subcellular distribution of HBx and CNP from the real line area of P3 panel.

Article Snippet: Membranes were blocked with 0.05% TBST (Tris-Buffered Saline Tween-20) containing 5% nonfat dry milk and probed with antibodies against CNP (Proteintech 1:3000), β-actin (Abcam, 1:5000), secondary antibodies conjugated to horseradish peroxidase were used and followed by ECL (Merck Millipore) detection.

Techniques: Infection, Staining, Control

Journal: BMC Pregnancy and Childbirth

Article Title: Maternal plasma pro-atrial and C-type natriuretic peptide levels and their associations with cardiovascular and renal function in the second half of normal pregnancy: a longitudinal study

doi: 10.1186/s12884-021-03824-3

Figure Lengend Snippet: Percentiles of maternal plasma C-type natriuretic peptide (pg/mL) at 22–40 weeks of gestational age (GA)

Article Snippet: The human C-type natriuretic peptide quantification kit (CSB-E08909h, CUSABIO, Lab-Tech AS, Norway) had an inter-assay CV of ≤10% and an intra-assay CV of ≤8%, and LOD was 15.6 pg/mL.

Techniques: Clinical Proteomics

Journal: Cell reports

Article Title: GluA2 overexpression in oligodendrocyte progenitors promotes postinjury oligodendrocyte regeneration

doi: 10.1016/j.celrep.2021.109147

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: The primary antibodies used in this study were rabbit anti-MBP (Cell Signaling Technology, Cat# 78896; 1:2,000), rabbit anti-CNPase (Phosphosolutions, Cat#325-CNP; 1:2,000), mouse anti-MAG (Santa Cruz Biotechnology, Cat# sc-166849; 1:4,000), mouse anti-MOG (Santa Cruz Biotechnology, Cat# sc-166172; 1:2,000), and mouse anti-β-actin (Santa Cruz Biotechnology, Cat# sc-47778; 1:2,000).

Techniques: Bicinchoninic Acid Protein Assay, SYBR Green Assay, Software, Microscopy, Imaging

Journal: American Journal of Physiology-Lung Cellular and Molecular Physiology

Article Title: C-type natriuretic peptide system in fetal ovine pulmonary vasculature

doi: 10.1152/ajplung.2001.281.2.l361

Figure Lengend Snippet: Fig. 4. Cumulative concentration-response curves to CNP in PA and PV isolated from late-gestation fetal lambs. All vessels were pre- treated with indomethacin, propranolol, and N-nitro-L-arginine (L-NNA). Relaxations are expressed as percentage of plateau norepi- nephrine (NE) constriction. n, No. of animals. PV relax significantly better than PA to CNP.

Article Snippet: The following pharmacological agents were used: indomethacin, DL-propranolol, N-nitro-L-arginine (L-NNA), L-norepinephrine hydrochloride, ANP (rat ANP 1–28), and CNP (human CNP 22; Peptides International, Louisville, KY).

Techniques: Concentration Assay, Isolation

Journal: American Journal of Physiology-Lung Cellular and Molecular Physiology

Article Title: C-type natriuretic peptide system in fetal ovine pulmonary vasculature

doi: 10.1152/ajplung.2001.281.2.l361

Figure Lengend Snippet: Fig. 7. Cumulative concentration-response curves to CNP in PA (A) and PV (B) isolated from late-gestation fetal lambs in the presence and absence of protein kinase G inhibitor, Rp-b-phenyl-1-N2-etheno- 8-bromoguanosine 39,59-cyclic monophosphorothionate (Rp-8-Br- PET-cGMPS, PET; 3 3 1025 M). All vessels were pretreated with indomethacin, propranolol, and L-NNA. Relaxations are expressed as percentage of plateau NE constriction. n, No. of animals. Pretreat- ment with Rp-8-Br-PET-cGMPS results in significant inhibition of CNP-mediated relaxation in PA and PV.

Article Snippet: The following pharmacological agents were used: indomethacin, DL-propranolol, N-nitro-L-arginine (L-NNA), L-norepinephrine hydrochloride, ANP (rat ANP 1–28), and CNP (human CNP 22; Peptides International, Louisville, KY).

Techniques: Concentration Assay, Isolation, Inhibition

Journal: American Journal of Physiology-Lung Cellular and Molecular Physiology

Article Title: C-type natriuretic peptide system in fetal ovine pulmonary vasculature

doi: 10.1152/ajplung.2001.281.2.l361

Figure Lengend Snippet: Fig. 6. Cumulative concentration-response curves to CNP in PA (A) and PV (B) isolated from late-gestation fetal lambs in the presence and absence of NPR-particulate guanylate cyclase inhibitor (HS- 142-1; 2 3 1025 M). All vessels were pretreated with indomethacin, propranolol, and L-NNA. Relaxations are expressed as percentage of plateau NE constriction. n, No. of animals. Pretreatment with HS- 142-1 results in significant inhibition of CNP-mediated relaxation in PA and PV.

Article Snippet: The following pharmacological agents were used: indomethacin, DL-propranolol, N-nitro-L-arginine (L-NNA), L-norepinephrine hydrochloride, ANP (rat ANP 1–28), and CNP (human CNP 22; Peptides International, Louisville, KY).

Techniques: Concentration Assay, Isolation, Inhibition

Journal: eLife

Article Title: A single-cell atlas of the cycling murine ovary

doi: 10.7554/elife.77239

Figure Lengend Snippet: Figure 6. Identification and validation of new secreted estrous staging markers. (A) Expression of granulosa cell transcripts varying by estrous cycle stage. (B) Validation of significantly up- and downregulated transcripts of secreted estrous staging markers by qPCR (n=5 per group, mean ± SEM, *p<0.05, **p<0.01, and ***p<0.005). (C) Localization of estrous staging markers by in situ hybridization (RNAscope) in ovarian sections. (D) Quantification of circulating estrous staging markers proteins in the blood by enzyme-Linked Immunosorbent assay (ELISA) (n=5 per group, mean ± SEM, *p<0.05, and ***p<0.005). (E) Summary of the timing of expression of estrous staging markers in the blood.

Article Snippet: Reagent type (species) or resource Designation Source or reference Identifiers Additional information Genetic reagent (Mus musculus) C57BL/6- Tg(UBC- GFP)30Scha/J Jackson Laboratory stock #004353 Antibody Smooth muscle alpha action (SMA) (Rabbit polyclonal) Abcam #5694 Dilution: 1:300 Commercial assay or kit ACTIVIN A commercial ELISA RnD systems #DAC00B Commercial assay or kit NPPC commercial ELISA Novus Bio #NBP2- 75790 Commercial assay or kit Tinagl1 commercial ELISA LS- Bio #LS- F49684 Commercial assay or kit PRSS35 commercial ELISA Mybiosource #MBS9717242 Commercial assay or kit RNA scope 2.5 HD Duplex detection kit ACD bio #322500 Commercial assay or kit RNA scope 2.5 HD red detection kit ACD bio #322360 Commercial assay or kit The target retrieval and protease plus reagents ACD bio #322330 Other Cdkn1a (M. musculus) NM_007669.4 ACD bio # 408551 RNAscope probe Other Cxcl14 (M. musculus) NM_019568.2 ACD bio #459741 RNAscope probe Morris, Meinsohn et al. eLife 2022;11:e77239.

Techniques: Biomarker Discovery, Expressing, In Situ Hybridization, RNAscope, Enzyme-linked Immunosorbent Assay