ppp control  (InvivoGen)

Journal: Molecular Cancer

Article Title: BRRIAR lncRNA alters breast cancer risk by modulating interferon signaling in cis and in trans

doi: 10.1186/s12943-025-02510-8

Figure Lengend Snippet: BRRIAR expression and immune activation in patient-derived samples. a High-resolution spatial transcriptomics map of one ER + breast tumor generated using the Curio Seeker platform. b Corresponding BRRIAR -positive cell states. c Spatial neighborhood analysis of BRRIAR -positive cells. Bars represent the proportion of neighbouring cell types within a defined radius. Error bars, SD ( n = 13 cells). p value was determined by Student’s t- test (* p < 0.05). d Scatter plot showing correlation between BRRIAR and RIG-I (left panel) or IRF7 (right panel) expression. Each black dot represents an individual cell at the co-expression peak. A linear regression line with its 95% confidence interval (blue shaded area) is overlaid. The correlation coefficient was calculated using Pearson correlation ( R ), and p values were determined using a two-sided t -test. e Spatially resolved BRRIAR expression across different tumor types using both 10x Visium and Curio Seeker platforms. BRRIAR -positive cells are depicted as colored dots. f-g Cytokine profiling for granzyme B, granzyme A, IFNγ and CCL5 secreted from HCMV-activated PBMCs (donors; D1-4) after f 48 h exposure to media harvested from T47D cells transfected with dsi-CON or dsi- BRRIAR then treated with 5’ppp-dsRNA for 3 h. The dsi-CON is a non-targeting control. Error bars, SEM ( n = 4). Or g 48 h exposure to media harvested from T47D cells transfected with IVT LacZ or IVT BRRIAR for 24 h. Error bars, SEM ( n = 4). p values were determined by two-way ANOVA with Sidak’s test (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). h Proposed model for BRRIAR as a positive regulator of RIG-I activation in ER + breast tumor cells to promote IFN production and support an anti-tumor immune response

Article Snippet: T47D cells were transfected with 3p-hpRNA (0.5 μg/ml; Invivogen) using Lipofectamine 3000, or 5’ppp-dsRNA (1 μg/ml) and its non-phosphorylated control dsRNA (Invivogen) using 6.25 μg of LyoVec transfection reagent (Invivogen) as per the manufacturer’s protocol.

Techniques: Expressing, Activation Assay, Derivative Assay, Generated, Transfection, Control

Journal: bioRxiv

Article Title: Human RIG-I deficiency confers susceptibility to Kaposi Sarcoma via loss of latency control

doi: 10.1101/2025.09.19.676307

Figure Lengend Snippet: A- LCLs from P1 and HC were stimulated for 24h with different RIG-I agonists and evaluated a panel of antiviral and antiproliferative interferon-stimulated gene (ISG) expression by QPCR. GAPDH is the housekeeping gene. B, C- A549-Dual™ KO-RIG-I cells were transfected with either an empty vector (EV), a wild-type RIG-I construct (RIG-I wt) or a RIG-I mutant construct (RIG-I Q393*). B- Immunoblotting was performed to evaluate RIG-I protein expression, and band intensities were quantified relative to GAPDH. C- Cells were then stimulated with 5’ppp or treated with IFNα (positive control) for 24h. Interferon regulatory factor (IRF) pathway activation was assessed by monitoring Lucia luciferase activity. Data are presented as relative luciferase units (RLU). Error bars represent standard deviation of the mean (SD) from triplicate experiments.

Article Snippet: 3p-hpRNA, 5’ppp-dsRNA, 5’ppp-dsRNA Control, Poly(dA:dT), Poly(I:C), R848, ODN 2006, Poly(dA:dT) and Poly(dG:dC) and Lyovec were from Invivogen.

Techniques: Expressing, Transfection, Plasmid Preparation, Construct, Mutagenesis, Western Blot, Positive Control, Activation Assay, Luciferase, Activity Assay, Standard Deviation