Journal: STAR Protocols

Article Title: Protocol for validating computationally predicted splice-altering variants using full-length gene reporter assays

doi: 10.1016/j.xpro.2026.104433

Figure Lengend Snippet: Transient splicing reporter assay using the genomic locus sequence reproduces the endogenous TREM2 and APOE isoform patterns (A) Schematic representation of the genomic TREM2 and APOE sequences cloned into the pcDNA3.1(+) vector. (B) Human cell lines expressing endogenous TREM2 (THP-1, HMC3), non-expressing HEK293T cells, and mouse microglial BV2 cells were transfected with the full-length (FL) TREM2 reporter. (C) HMC3 and HEK293T cells expressing endogenous APOE were transfected with the FL APOE reporter. HB, endogenous TREM2 (B) and APOE (C) transcripts from human brain (frontal cortex). cDNA synthesis was primed with either oligo(dT) or a plasmid-specific RT primer (P) positioned downstream of the gene insert. Arrows mark RT-PCR primer positions. Amplified products were resolved by agarose gel electrophoresis.

Article Snippet: HEK293T , ATCC , CRL-3216.

Techniques: Reporter Assay, Sequencing, Clone Assay, Plasmid Preparation, Expressing, Transfection, cDNA Synthesis, Reverse Transcription Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis

Journal: eLife

Article Title: Interplay between PML NBs and HIRA for H3.3 dynamics following type I interferon stimulus

doi: 10.7554/eLife.80156

Figure Lengend Snippet:

Article Snippet: Human BJ primary foreskin fibroblasts (ATCC, CRL-2522), human IMR90 fetal lung fibroblasts (ATCC, CCL-186), human HEK 293T embryonic kidney cells (Intercell, AG) and mouse MEFs embryonic fibroblasts Pml +/+ or Pml -/- (from Dr. Lallemand-Breitenbach, and whose cell identity was authenticated by STR profiling) were cultivated in DMEM medium (Sigma-Aldrich, D6429) containing 10% of fetal calf serum (FCS) (Sigma-Aldrich, F7524), 1% of penicillin/streptomycin (Sigma-Aldrich, P4458), at 37 °C under 5% CO2 and humid atmosphere.

Techniques: Transduction, Recombinant, Plasmid Preparation, Clone Assay, Mutagenesis, Sequencing, Quantitative RT-PCR, Concentration Assay, In Situ, Software, ChIP-sequencing