solution rg7834  (MedKoo Inc)

Journal: PLOS Pathogens

Article Title: Host species-specific mutations in the thumb domain of the 3D pol polymerase are required for efficient replication of human hepatitis A virus in mice

doi: 10.1371/journal.ppat.1014213

Figure Lengend Snippet: (A) HAV RNA abundance in lysates of human (Huh-7.5) versus murine (AML12) hepatocytes 5-7 days following transfection of in vitro transcribed p16 (3D pol -R468), p16/3D pol -K468, or p16/3D pol -G473 RNA. Data are mean values from 3 independent experiments, each with 2-3 technical replicates. The dashed lines indicate mean HAV RNA abundance in each cell type (n = 3) at harvest when treated with the potent antiviral RG7834 (200nM) immediately after transfection. (B) Percentage of HAV transcripts with the 3D pol -R468 allele determined by high-throughput RNA sequencing of RT-PCR amplimers from cultures of Huh-7.5 and AML12 cells (two independent cultures of each) following transfection of a nominal 50:50 mixture of p16 (3D pol -R468) and p16/3D pol -K468 RNA. Read counts ranged from 7820 in the inoculum mix, to a mean of 7817 at day 14 in Huh-7.5 cells and 620 at 10 days in AML12 cells. HAV RNA abundance was insufficient for quantitation at day 14 in AML12 cells. p-values by Fisher’s exact test; p = 0.009 for cell type-specific differences between days 0-10 by two-way repeated measures ANOVA.

Article Snippet: The S-isomer of the antiviral compound RG7834 was purchased from MedChemExpress (#HY-117650A), and used to assess HAV replication at a concentration of 200nM.

Techniques: Transfection, In Vitro, High Throughput Screening Assay, RNA Sequencing, Reverse Transcription Polymerase Chain Reaction, Quantitation Assay

Journal: Microbiology Spectrum

Article Title: A visible assay for evaluating the inhibitory activity of drug and antibody against HBV infection

doi: 10.1128/spectrum.02638-25

Figure Lengend Snippet: Evaluating the inhibitory efficacy of RG7834 drugs against HBV infection by immunospot assay. HepG2-NTCP cells were infected with 300 HBV gec/cell at 37°C for 24 h, and then incubated with RG7834 drugs. After 7 days, the individual HBV-infected cell was identified by HRP-conjugated cAbD4 ( A ) and calculated at different drug concentrations ( B ). ( C ) Inhibition curves of RG7834 against HBV infection from ( B ) were depicted. ( D ) HBeAg in supernatant from day 3 to 6 was detected by ELISA. ( E ) Inhibition curves of RG7834 against HBV infection from ( D ) were depicted. The dashed line indicates a 50% reduction in viral infectivity. The experiment was independently performed at least twice, and one representative result was shown.

Article Snippet: Myrcludex B lipopeptides (MGTNLSVPNPLGFFPDHQLDPAFGANSNNPDW-DFNPNKDHWPEANKVG), RG7834, and NVR 3-778 were purchased from MedChemExpress, and the control peptide (DTDFVNEFYAYLRKH) was synthesized by GenScript Biotech Corporation.

Techniques: Infection, Incubation, Inhibition, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: CDK12/13 inhibition disrupts nucleolar morphology and promotes aberrant expression of IGS transcripts

doi: 10.1101/2025.03.01.640972

Figure Lengend Snippet: (A) Heatmap showing the differential expression levels of RNA exosome and nuclear exosome co-factors in IMR32, Kelly, MG63.3, and 143B cells treated with THZ531 (500 nM x 6 h) or E9 (200 nM x 6 h) compared to DMSO treated control. Expression levels were obtained from public RNA-seq datasets GSE113314 and GSE132233 from the GEO database. Differential expression is represented as log2(fold change), with upregulation shown in red and downregulation in blue. (B) Violin plot showing the average gene length of core RNA exosome, NEXT, PAXT and TRAMP components. (C) IF analysis of poly(A) and 28S rRNA in OS cells transfected with siRNA against ZCCHC7 or TENT4A mRNA and incubated with E9 (200 nM x 6 h). Scale bar, 5 μm, inset, 1 μm. (D) IF analysis of the same markers as in (C) in OS cells treated with E9 (200 nM x 6 h) and/or RG-7834 (10 μM x 6 h). Scale bar, 5 μm, inset, 1 μm. (E) RIP-qPCR analysis of MTREX binding to 18S, 28S, H16 and H27 rRNAs in OS cells treated with E9 (200 nM x 6 h). Data represent mean ± S.D. of 2 independent replicates. RNA enrichment was calculated as (% of input of rRNA or IGS)/(% of input of GAPDH). (F) IF analysis of 28S rRNA and MTREX in OS cells treated as in (E). Scale bar, 5 μm. (G) RIP-qPCR analysis of EXOSC2 binding to 18S, 28S, H16 and H27 rRNAs in OS cells treated as in (E). Data represent mean ± S.D. of 2 independent replicates. RNA enrichment was calculated as in (E). (H) Schematic model for pAR rings accumulation following CDK12/13 inhibition in OS cells.

Article Snippet: E9, THZ-531, BSJ-4-116, BSJ-4-23, BSJ-4-23NC, THZ1 and YKL-5-124 were synthesized and provided by Dr. Nathanael Gray (Dana-Farber Cancer Institute, Stanford University). dTAG13 (Cat #: 66-055) and Actinomycin D (Cat #: 11805017) were purchased from Fisher Scientific, CR8 (Cat #: C3249) and CX-5461 (Cat #: 5.09265) from Sigma Aldrich, NVP-2 (Cat #: HY-12214A) and RG-7834 (Cat #: HY117650A) from MedChem Express, and flavopiridol from Selleck Chemicals (Cat #: S2679).

Techniques: Quantitative Proteomics, Control, Expressing, RNA Sequencing, Transfection, Incubation, Binding Assay, Inhibition

Journal: bioRxiv

Article Title: CDK12/13 inhibition disrupts nucleolar morphology and promotes aberrant expression of IGS transcripts

doi: 10.1101/2025.03.01.640972

Figure Lengend Snippet: (A) 3’-Poly(A) RNA-sequencing tracks of ZCCHC14, MTREX and ZCCHC7 genes in IMR32 and Kelly cells treated with THZ531 (400 nM x 6 h) and DMSO. Intronic polyadenylation sites are marked with *. (B) IF analysis of poly(A) and 28S rRNA in OS cells transfected with siRNA against ZCCHC7 or TENT4A mRNA. Scale bar, 5 μm. (C) RT-qPCR analysis of 18S (H4), 28S (H13) and H16 region in OS cells treated with RG-7834 or transfected with siRNA against TENT4A mRNA. (D) Cell viability in OS cells treated or transfected as in (C). Data represent mean ± S.D. of 3 independent replicates.

Article Snippet: E9, THZ-531, BSJ-4-116, BSJ-4-23, BSJ-4-23NC, THZ1 and YKL-5-124 were synthesized and provided by Dr. Nathanael Gray (Dana-Farber Cancer Institute, Stanford University). dTAG13 (Cat #: 66-055) and Actinomycin D (Cat #: 11805017) were purchased from Fisher Scientific, CR8 (Cat #: C3249) and CX-5461 (Cat #: 5.09265) from Sigma Aldrich, NVP-2 (Cat #: HY-12214A) and RG-7834 (Cat #: HY117650A) from MedChem Express, and flavopiridol from Selleck Chemicals (Cat #: S2679).

Techniques: RNA Sequencing, Transfection, Quantitative RT-PCR