Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Regulation of Aldosterone Secretion by Substance P and the Neurokinin Type 1 Receptor in Aldosterone‐Producing Adenomas

doi: 10.1161/JAHA.125.045539

Figure Lengend Snippet: A , Quantitative RT‐PCR analysis of TACR1 , TACR2 , and TACR3 mRNAs in 42 independent APAs. Each dot indicates 1 adenoma. Data are presented as mean±SEM. P =0.012; P <0.0001. B , Comparison of the expression levels of the TACR1 gene between different APA subgroups. classical APAs; nonclassical APAs (left panel) P =0.045; NKD and KCNJ5 (middle panel); CYP11B2‐positive nodules in nonclassical APAs (<2 nodules vs ≥2 nodules, right panel). C through F , Distribution of NK1R immunostaining in APA and adjacent adrenal tissue representative of n=56 independent APAs. C, E, F, NK1R staining in a classical APA and its adjacent adrenal cortex. C , Overall NK1R staining pattern in an APA and the adjacent adrenal cortex. D , Closer view of the NK1R staining in a different classical APA, highlighting areas of staining present in the zona glomerulosa or organized in micronodules within the adrenal tissue adjacent to the adenoma (arrows). E , Close‐up view of NK1R‐positive staining in the ZG and nerve ganglia in the adrenal cortex adjacent to the adenoma). F , High magnification view illustrating the distribution of NK1R staining in a group of adenoma cells. Arrow and arrowhead show membrane and cytoplasmic NK1R staining, respectively. APA indicates aldosterone‐producing adenoma; C, classical; Ca, capsule; G, nerve ganglia; KCNJ5 , KCNJ5 mutation detected; NC, nonclassical; NKD, no KCNJ5 mutation detected; NK1R, neurokinin type 1 receptor; RT‐PCR, reverse transcription polymerase chain reaction; and ZG, zona glomerulosa.

Article Snippet: Each sample was analyzed in duplicates, and cDNA quantification was normalized to PPIA (cyclophilin) using the ΔCt method and standard curves generated from polyA mRNAs or TACR1 human quantitative PCR template ( HK210362 , Origene, Rockville, USA).

Techniques: Quantitative RT-PCR, Comparison, Expressing, Immunostaining, Staining, Membrane, Mutagenesis, Reverse Transcription Polymerase Chain Reaction, Reverse Transcription, Polymerase Chain Reaction

Journal: Nature Communications

Article Title: The neuropeptide substance P regulates aldosterone secretion in human adrenals

doi: 10.1038/s41467-020-16470-8

Figure Lengend Snippet: a Representative western blots showing the kinetics of the phospho-ERK (upper panels) and ERK (lower panel) levels in response to SP (from 10 −6 to 10 −10 M) and Ang II (10 −6 M) after incubation of cultured cells for 5, 15, 30 and 45 min (three blots processed in parallel showing samples derived from the same culture experiment). Immunolabellings for phospho-ERK and ERK were successively performed on the same membranes. Ang II and vinculin were used as positive and loading controls, respectively. b Time course of ERK phosphorylation normalised to ERK in response to SP (10 −7 M). Data are normalised to basal level (0 min) of each repetitive experiment. Data are presented as mean ± SEM of three independent cultures. c Representative western blots showing the effect of increasing doses of SP administrated for 5 min on phospho-ERK (left panel) and ERK (right panel). d Dose-response curves of SP-induced ERK phosphorylation normalised to ERK for 5, 15 and 30 min of incubation (T5, T15 and T30), illustrated in c and Supplementary Fig. . Data are expressed as percentage basal level (Bl). e Effect of the ERK pathway inhibitor PD0325901 (10 –8 M) on SP-induced aldosterone secretion. Data are expressed as median ± IQR, minima and maxima ( n = 4 cultures). They were analysed by two-tailed Mann–Whitney test ( p = 0.028 for SP versus SP + PD0325901). f Effect of SP (10 −6 M, 24 h) on the expression levels of genes encoding cholesterol transporter and steroidogenic enzymes. mRNA expression levels were normalised to PPIA. Data are expressed as median ± IQR, minima and maxima ( n = 4, 8, 8 and 11 independent cultures for STAR , HSD3B2 , CYP21A2 and CYP11B2 , respectively). They were analysed by two-tailed Mann–Whitney test ( p = 0.48 for STAR , p = 0.02 for HSD3B2 , p = 0.0002 for CYP21A2 and p = 0.14 for CYP11B2 . * p < 0.05; *** p < 0.001.

Article Snippet: Quantification of cDNAs was normalised to PPIA (cyclophilin) by using standard curves established by using dilution series of cDNAs derived from polyA mRNAs or TACR1 human qPCR Template (HK210362, Origene).

Techniques: Western Blot, Incubation, Cell Culture, Derivative Assay, Phospho-proteomics, Two Tailed Test, MANN-WHITNEY, Expressing

Journal: Oncology reports

Article Title: Cholesterol synthesis inhibitor RO 48-8071 suppresses transcriptional activity of human estrogen and androgen receptor.

doi: 10.3892/or.2014.3332

Figure Lengend Snippet: Figure 1. Relative OSC mRNA expression in breast cancer at different stages of growth. Human qPCR-ready TissueScan™ cDNA Arrays (cat. no. TSRT101) were obtained from OriGene and real-time PCR was conducted as described in Materials and methods. Results showed variable levels of expression of OSC mRNA within different tumors compared with corresponding normal tissue. OSC, oxidosqualene cyclase.

Article Snippet: Human tissue qPCR. qPCR-ready TissueScanTM cdNA Array (OriGene starter kit, cat. no. TSRT101) was initially used to determine the expression of OSC in human breast cancer tissues following guidelines recommended by the manufacturer.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Oncology reports

Article Title: Cholesterol synthesis inhibitor RO 48-8071 suppresses transcriptional activity of human estrogen and androgen receptor.

doi: 10.3892/or.2014.3332

Figure Lengend Snippet: Figure 2. Relative OSC mRNA expression in distinct breast cancer tissues at different stages of growth (stage I-III). Human qPCR-ready TissueScan™ cDNA Arrays (cat. no. BCRT101) were obtained from OriGene and real‑time PCR was conducted as described in Materials and methods. Results showed no significant difference in OSC mRNA expression between hormone-dependent (ER+, PR+, HER2+) and triple negative breast cancer at various stages of growth. *Triple negative breast tumor, **ER/PR-negative/Her-2neu high tumors; remaining bars represent ER/PR positive tumors. OSC, oxidosqualene cyclase.

Article Snippet: Human tissue qPCR. qPCR-ready TissueScanTM cdNA Array (OriGene starter kit, cat. no. TSRT101) was initially used to determine the expression of OSC in human breast cancer tissues following guidelines recommended by the manufacturer.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: bioRxiv

Article Title: Slow scission of single synaptic vesicles by Dynamin at physiological temperature

doi: 10.1101/2025.03.18.643914

Figure Lengend Snippet: a, Normalized average fluorescent traces of Dynamin I-EGFP overexpressed in Xenapses responding to a 50 AP 40 Hz stimulus at RT (black) and 37°C (red). mean ± s.e.m. of n = 19 measurements at RT vs n = 27 measurements at 37°C both from 5 biological replicates. b, Normalized average trace of single Dynamin events elicited with 1 or 2 AP. mean ± s.e.m. c, Normalized cumulative event duration distribution (pORANGE Dynamin I-EGFP knock-in) for RT (red) and 37°C (black). For 37°C, 1031 events (evoked with 1 or 2 AP) consisting of n = 16 measurements from 9 biological replicates. For RT, 119 events (evoked with 2 or 5 AP) consisting of n = 10 measurements from 2 biological replicates.

Article Snippet: These were then individually ligated into BbsI digested pORANGE cloning template vector (Addgene Plasmid #131471) enabled by the duplex having CACCG and AAAC overhangs at either end.

Techniques: Knock-In

Journal: bioRxiv

Article Title: Slow scission of single synaptic vesicles by Dynamin at physiological temperature

doi: 10.1101/2025.03.18.643914

Figure Lengend Snippet: a, Schematic depicting EGFP targetting to the Dynamin I gene by CRISPR-Cas9 based pORANGE strategy (BSE : Bundle signalling element, PRD : Proline rich domain). b, TIRF-microscopy image 3 s after 1 AP stimulus of Xenapses expressing EGFP tagged to endogenous Dynamin I with the Dynamin puncta circled in red; scale bar, 3 µm. c, Representative traces of individual evoked Dynamin recruitment and scission events occurring at various time points after the stimulus. d, An exemplar trace showing the stepwise Dynamin recruitment and annotated with measurable properties. Fit (red) obtained by applying the Autostepfinder step-detection function. Shown below are fluorescence images of the single Dynamin scission event at times indicated; scale bar, 0.5 µm. e, Event duration distribution, f, Event latency distribution and g , Event time to scission distribution. e,f,g are from the same data set of 1129 events in response to 1 or 2 AP and analyzed from n = 16 measurements from 7 biological replicates.

Article Snippet: These were then individually ligated into BbsI digested pORANGE cloning template vector (Addgene Plasmid #131471) enabled by the duplex having CACCG and AAAC overhangs at either end.

Techniques: CRISPR, Microscopy, Expressing, Fluorescence

Journal: bioRxiv

Article Title: Slow scission of single synaptic vesicles by Dynamin at physiological temperature

doi: 10.1101/2025.03.18.643914

Figure Lengend Snippet: Representative traces of pre-stimulus spontaneous (a) and straddle (b) Dynamin recruitment and scission events. c, d, Average trace of spontaneous single Dynamin events centered in space and time along the scission frame and the corresponding average trace of Clathrin fluorescence (d) . Inset images show normalized average fluorescence at time points indicated. mean ± s.e.m. of 38 events from n = 7 measurements with both sets of data from 3 biological replicates. e, Normalized cumulative event duration distributions for Dynamin I EGFP knock-in by pORANGE for evoked (black) and spontaneous events (red, including straddle events). 1031 evoked and 190 spontaneous events both from same data set consisting of n = 16 measurements and 9 biological replicates. f, Normalized inverse cumulative distribution of scission waiting times (time course of endocytosis) for all Dynamin scission events (solid red) and excluding straddle events (dashed red). A monoexponential fit (black) to the decaying phase after 4 seconds (median duration time) yields a time constant of 9.64 seconds. 206 evoked (1 AP) events and 27 straddle events both from n = 4 measurements and biological replicates.

Article Snippet: These were then individually ligated into BbsI digested pORANGE cloning template vector (Addgene Plasmid #131471) enabled by the duplex having CACCG and AAAC overhangs at either end.

Techniques: Fluorescence, Knock-In

Journal: bioRxiv

Article Title: Slow scission of single synaptic vesicles by Dynamin at physiological temperature

doi: 10.1101/2025.03.18.643914

Figure Lengend Snippet: a, Schematic depicting EGFP insertion to the Dynamin I 3‘UTR by CRISPR-Cas9 based TKIT strategy. b, Normalized cumulative stimulus-evoked event duration for Dynamin I EGFP-tagged by pORANGE (black) and TKIT(red) knock-in strategies. 118 events from n = 6 measurements and 2 biological replicates with post-stimulus events evoked at 2 or 5 AP. c, Representative TIRFM image of a Xenapse formed on a 2.5 µm micropattern expressing pORANGE tagged Dynamin-EGFP. Several puncta of Dynamin recruitment are visible 1 s after 2 AP; scale bar, 1 µm. d, Normalized cumulative event durations including straddle events for Xenapses growing on 5 µm (black) and 2.5 µm (red) micropatterns. e , Normalized inverse cumulative distribution of waiting times of scission (time course of endocytosis) for Xenapses growing on 5 µm (black) and 2.5 µm (red) micropatterns. For pORANGE (b) and 5 µm spots (d,e) , same data set as for post-stimulus in Fig.3f . and for 2.5 µm spots (d,e) , 65 events from n = 7 measurements and 2 biological replicates with post-stimulus events evoked at 1, 2 or 5 AP.

Article Snippet: These were then individually ligated into BbsI digested pORANGE cloning template vector (Addgene Plasmid #131471) enabled by the duplex having CACCG and AAAC overhangs at either end.

Techniques: CRISPR, Knock-In, Expressing

Journal: bioRxiv

Article Title: Slow scission of single synaptic vesicles by Dynamin at physiological temperature

doi: 10.1101/2025.03.18.643914

Figure Lengend Snippet: a, Event latency distribution for Dynamin I-EGFP (pORANGE knock-in) scission events in response to 1 (n = 206), 2 (n = 208) and 5 AP (n = 233). n = 4 measurement from 3 biological replicates for 1 AP and n = 6 measurements from 5 biological replicates for 2 and 5 AP. b , SEM images of the two major observed classes of Clathrin coated structures, domed and pit, and the transition between the two. c , Density of domed (8.03 ± 3.62 per mm 2 ) and pit (0.81 ± 0.36 per mm 2 ) structures from SEM images of unroofed Xenapses (n = 17).

Article Snippet: These were then individually ligated into BbsI digested pORANGE cloning template vector (Addgene Plasmid #131471) enabled by the duplex having CACCG and AAAC overhangs at either end.

Techniques: Knock-In

Journal: Protein science : a publication of the Protein Society

Article Title: Functional studies of N-terminally modified CYP2J2 epoxygenase in model lipid bilayers.

doi: 10.1002/pro.2280

Figure Lengend Snippet: Figure 1. Design and expression of CYP2J2 mutants. (A) CYP2J2 wild-type and design of N-terminal mutant constructs. Boxed residues denote substitution of native Leu with Ala (GCT). Bolded residues represent introduction of AT-rich silent mutations. Underlined residues signify introduction of a hydrophilic motif. Gaps represent deletion of N-terminal amino acids. (B) CYP2J2 N-terminal mutant expression and solubility in DH5alpha cells. The bar graph denotes expression yields that were highly dependent on N-terminal modifications. The line plot signifies percentage of protein found in the soluble fraction during purifica- tion, which largely corresponded to N-terminal truncations.

Article Snippet: The native human CYP2J2 gene was obtained from OriGene.

Techniques: Expressing, Mutagenesis, Construct, Solubility

Journal: Protein science : a publication of the Protein Society

Article Title: Functional studies of N-terminally modified CYP2J2 epoxygenase in model lipid bilayers.

doi: 10.1002/pro.2280

Figure Lengend Snippet: Figure 2. Characterization of CYP2J2 constructs. (A) CYP2J2 mutants were incorporated into Nanodiscs and purified using SEC-HPLC. SDS-PAGE was used to verify successful isolation of each respective CYP2J2-ND. The first column contains a protein ladder standard showing that the molecular weight of M2D19HG is close to the 50–60 kDa standard and the scaffold- ing protein MSP1T2 has molecular weight close to 24 kDa. The exact molecular weights were confirmed using ESI-MS. (B) CO-binding spectra were used to characterize each construct and gauge the protein folding after expression and purifica- tion. Each construct exhibited 100% P450 content, which is indicative of a well-folded and functional CYP. (C) SEC-HPLC was applied to characterize CYP2J2 mutant oligomerization states in phosphate buffer (0.1M). The CYP2J2 elution profile was produced by monitoring the heme absorbance at 417 nm. Using water-soluble protein standards, we observed that the constructs smaller oligomeric state corresponded closely with the size of the N-terminal truncation and followed M2SMG M2D5HG > M2D19HG > M2D34G.

Article Snippet: The native human CYP2J2 gene was obtained from OriGene.

Techniques: Construct, SDS Page, Isolation, Molecular Weight, Binding Assay, Expressing, Functional Assay, Mutagenesis, Produced

Journal: Protein science : a publication of the Protein Society

Article Title: Functional studies of N-terminally modified CYP2J2 epoxygenase in model lipid bilayers.

doi: 10.1002/pro.2280

Figure Lengend Snippet: Figure 4. Incorporation of CYP2J2 constructs in Nanodiscs and Nanodisc stability assay. (A) Visual molecular dynamics (VMD) was used to construct a representative model of CYP2J2 incorporated into Nanodiscs (CYP2J2 model was obtained using Phyre). In the model, blue represents the membrane scaffolding protein (MSP1D1), the gray are the phospholipids, and the protein ribbon model is the CYP2J2 enzyme. (B) Membrane association of M2D34HG model. Highlighted residues in the A helical region (Trp48, Arg49, Leu50, Pro51, Phe52, Leu53, Phe57, Leu58, and Phe61), Lys122 in B0 helix and in the FG loop region (Trp235, Phe239, and Leu240) are predicted by OMP software to be associated with the lipid membrane. Predicted Gibbs free energy for this interaction is 13.8 kcal mol21. (C) CYP2J2 constructs were incorporated into Nanodiscs and monitored at 417 nm SEC-HPLC. The single peak corresponds to homogeneous CYP2J2-Nanodiscs. (D) The relative stability of each CYP2J2 construct incorporated into Nano- discs was monitored using SEC-HPLC over a 7-day period at 4C. The percent area of the 417-nm peak corresponding to heme absorption was monitored at the established CYP2J2-Nanodisc elution time (26 min). We observed a similar relative stability for each of the constructs with an approximate 10–20% degradation over the time course.

Article Snippet: The native human CYP2J2 gene was obtained from OriGene.

Techniques: Construct, Stability Assay, Membrane, Scaffolding, Software

Journal: Protein science : a publication of the Protein Society

Article Title: Functional studies of N-terminally modified CYP2J2 epoxygenase in model lipid bilayers.

doi: 10.1002/pro.2280

Figure Lengend Snippet: Figure 5. Schematic of NADPH oxidation and ebastine hydroxylation by a CPR-CYP2J2 Nanodisc complex. Electrons are shuttled from NADPH to CYP2J2 via cytochrome P450 reductase redox centers (step 1). These electrons subsequently drive CYP2J2 P450 catalysis of ebastine to hydroxyebastine (step 2). Coupling efficiency is measured as the ratio of ebastine hydroxylation (step 2) over NADPH oxidation (step 1). [Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]

Article Snippet: The native human CYP2J2 gene was obtained from OriGene.

Techniques:

Journal: bioRxiv

Article Title: Multiplex labeling and manipulation of endogenous neuronal proteins using sequential CRISPR/Cas9 gene editing

doi: 10.1101/2022.01.02.474730

Figure Lengend Snippet: A. Illustration of the problem of multiplex knock-in strategies based on NHEJ. With simultaneous editing of multiple genes, donor DNAs can be integrated in either allele, leading to crosstalk. B. Proposed solution for multiplex knock-ins. By introducing a delay between two genome editing events, crosstalk can be avoided. CAKE is designed to control the delay between the two events, using Cre- or Flp-recombinase. C. CAKE strategy. The Cre OFF vector is active in the absence of Cre, leading to editing of the first target gene, and removal of the Cre OFF donor for subsequent genomic integration. Upon addition of Cre, gRNA expression from Cre OFF is deactivated, and gRNA expression from Cre ON vector is enabled for editing the second target gene. All donors contain a fluorophore or epitope tag flanked by a PAM- and target sequence (not shown, see and Willems et al ., 2020). D. Example confocal image of a Cre OFF β3-tubulin-GFP and Cre ON Halo-β-actin knock-in. 20 µL lenti-Cre was added at DIV 7, and cells were fixed at DIV 14.

Article Snippet: The Cre ON knock-in vector (pOC1) is based on pORANGE LOX from ( Willems et al ., 2020 , Addgene #139651), where CAG HA-SpCas9 was removed with XmaJI and NotI and replaced with primers AJ19164 and AJ19165 using primer ligation.

Techniques: Multiplex Assay, Knock-In, Control, Plasmid Preparation, Expressing, Sequencing

Journal: bioRxiv

Article Title: Multiplex labeling and manipulation of endogenous neuronal proteins using sequential CRISPR/Cas9 gene editing

doi: 10.1101/2022.01.02.474730

Figure Lengend Snippet: A. Overview of original CAKE knock-in mechanism introduced in ( Willems et al ., 2020 ), referred to as 2A-Cre throughout this Figure. The constitutively active knock-in labels β3-tubulin with GFP-2A-Cre. Cre expression from this allele then activates gRNA expression for the Cre ON Halo-β-actin knock-in. B. Overview of improved CAKE constructs introduced in this study (see main text) which will be referred to as Lenti-Cre throughout this Figure. Lentivirus encoding for Cre is added at DIV 7, which switches off gRNA expression for Cre OFF , and switches on gRNA expression for Cre ON . C. Example confocal images of CAKE double knock-ins using the 2A-Cre or Lenti-Cre mechanism. D. Average number of single and double knock-in cells per coverslip for 2A-Cre and Lenti-Cre CAKE. 2A-Cre n = 8 coverslips, lenti-Cre n = 7 coverslips, N = 4 independent cultures. β3-tubulin-GFP p = 0.50, Halo-β-actin p = 0.29, double knock-in p = 0.53, incorrect knock-in * p = 0.019, unpaired t -test. E. Average fluorescence intensity in proximal dendrites of single knock-ins, normalized per culture. For β3-tubulin-GFP: 2A-Cre n = 21 cells, lenti-Cre n = 25 cells, N = 3 independent cultures. For Halo-β-actin: 2A-Cre n = 18 cells, lenti-Cre n = 14 cells, N = 3 independent cultures. β3-tubulin-GFP * p = 0.001, Halo-β-actin p = 0.29, unpaired t -test. F. Example confocal image of an incorrect double knock-in. Fluorescence signal of both GFP and Halo are consistent with β3-tubulin distribution, suggesting that the Halo donor was inserted in Tubb3 .

Article Snippet: The Cre ON knock-in vector (pOC1) is based on pORANGE LOX from ( Willems et al ., 2020 , Addgene #139651), where CAG HA-SpCas9 was removed with XmaJI and NotI and replaced with primers AJ19164 and AJ19165 using primer ligation.

Techniques: Knock-In, Expressing, Construct, Fluorescence

Journal: bioRxiv

Article Title: Multiplex labeling and manipulation of endogenous neuronal proteins using sequential CRISPR/Cas9 gene editing

doi: 10.1101/2022.01.02.474730

Figure Lengend Snippet: A. Overview of pORANGE ( Willems et al ., 2020 ) and the CAKE template vectors introduced in this study. All vectors contain a multiple cloning site for addition of the donor DNA. B. Cloning guide to create CAKE knock-in constructs. The example shows a step-by-step protocol for pOC2 Cre OFF knock-in vectors, but identical cloning steps apply to all pOC vectors shown in A. #1 shows the empty pOC2 template vector. After digestion with BbsI, a primer pair encoding for the gRNA target sequence is added to obtain the intermediate construct (#2). The donor DNA, which always contains a protospacer adjacent motif (PAM) and target sequence ( Suzuki et al ., 2016 ) is obtained with standard PCR techniques. of Willems et al ., (2020) contains a detailed description on design of the donor DNA. The donor is cloned using HindIII and MluI restriction sites to obtain the final pOC2 knock-in construct (#3). Most of the knock-in constructs used in this study contain BmtI and AfeI restriction sites around the fluorophore, for universal exchange of donors between knock-in vectors, without the need of PCR (shown in #4, optional). The entire knock-in cassette, containing U6-driven gRNA expression, Lox551 and/or Frt sites and the donor DNA, can be removed from pOC vectors using PscI and MluI restriction sites.

Article Snippet: The Cre ON knock-in vector (pOC1) is based on pORANGE LOX from ( Willems et al ., 2020 , Addgene #139651), where CAG HA-SpCas9 was removed with XmaJI and NotI and replaced with primers AJ19164 and AJ19165 using primer ligation.

Techniques: Cloning, Knock-In, Construct, Plasmid Preparation, Sequencing, Clone Assay, Expressing

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Hepatitis B Virus Capsid Assembly Modulators, but Not Nucleoside Analogs, Inhibit the Production of Extracellular Pregenomic RNA and Spliced RNA Variants

doi: 10.1128/AAC.00680-17

Figure Lengend Snippet: Effect of CAMs and nucleos(t)ide analogs on intracellular HBV RNA and DNA. (A and B) Uninfected and infected HepaRG cells were treated with DMSO or 30 μM BAY 41-4109_IE, BAY 41-4109, LMV, TFV, or NVR 3-1983 for 6 days prior to Northern blot analysis of intracellular ribosomal 18S and 28S RNA (A, top), HBV pgRNA and preS/S mRNA transcripts (A, middle), and encapsidated pgRNA (A, bottom) and Southern blot analysis of intracellular encapsidated HBV DNA (B). (C to F) Infected HepaRG cells were treated with increasing concentrations of NVR 3-1983 or LMV for 6 days, prior to measuring the amonts of intracellular HBV DNA (C, E) or encapsidated HBV RNA (●), total HBV mRNA transcripts (■), and beta-actin RNA (▲) (D, F). (G and H) Effect of 10 μM LMV or BAY 41-4109 on HBV cccDNA (G) and secreted HBsAg (H). Data points and error bars represent means and standard deviations from at least three independent studies, respectively.

Article Snippet: Plasmids containing either the genotype D HBV genome or the human beta-globin gene (Origene) were used to establish standard curves.

Techniques: Infection, Northern Blot, Southern Blot