Journal: Molecular Cell

Article Title: POINT technology illuminates the processing of polymerase-associated intact nascent transcripts

doi: 10.1016/j.molcel.2021.02.034

Figure Lengend Snippet:

Article Snippet: The isolated RNA was fragmented to 150-200 nt at 90°C for 10 min according to a protocol of NEBNext Ultra II Directional RNA library prep kit to prepare PCR library.

Techniques: Sequencing, cDNA Library Assay, Software

Journal: The Journal of Biological Chemistry

Article Title: Vitamin D signaling inhibits HBV activity by directly targeting the HBV core promoter

doi: 10.1016/j.jbc.2021.101233

Figure Lengend Snippet: Calcitriol inhibits HBV core promoter transcripts, HBV secretory proteins, but not HBV virion secretion. The 1.3×-HBV genome construct was transfected in HepG2 and Huh7 cells, whereas HepG2.2.15 stably expresses HBV genome. In the HBV infection model, HBV particles derived from HepG2.2.15 cells were used to infect HepG2-NTCP cells as described in the section. Vehicle control (without calcitriol) or 10 nM calcitriol was added immediately after transfection or infection, and samples were processed as described in the section. RNA isolation and quantitation by quantitative PCR (qPCR) shows that ( A ) HBV pcRNA and ( B ) HBV pgRNA are suppressed in the presence of calcitriol. Quantitation of HBV secretory proteins by ELISA demonstrates that calcitriol negatively regulates ( C ) HBeAg and ( D ) HBsAg. E , finally, HBV virion secreted in supernatant was captured 72 h after ligand treatment, and its DNA was quantitated by qPCR. The virion secretion in the presence of calcitriol was marginally reduced in all HBV cell culture models (up to 12%) in vitro , though the observation was not statistically significant in any model. All data are means ± SD for three independent experiments (n = 3). ∗ p < 0.05, ∗∗ p < 0.01, and NS (analyzed by paired Student's t test). HBeAg, hepatitis B “e” antigen; HBsAg, hepatitis B surface antigen; HBV, hepatitis B virus; NS, not significant; NTCP, sodium taurocholate cotransporting polypeptide; pcRNA, precore RNA; pgRNA, pregenomic RNA.

Article Snippet: About 150 μl of the supernatant was added to each well of the HBsAg ELISA plate, along with DNaseI (New England Biolabs) and incubated for 2 h at 37 °C for immunocapture of the viral particles.

Techniques: Construct, Transfection, Stable Transfection, Infection, Derivative Assay, Isolation, Quantitation Assay, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Cell Culture, In Vitro