Journal: bioRxiv

Article Title: Structure of zebrafish NLRP3 reveals a novel mode of inflammasome activation

doi: 10.64898/2026.04.17.719140

Figure Lengend Snippet: a , LDH release assay in NLRP3-KO BlaER1 cells reconstituted with the indicated FLAG-tagged NLRP3 constructs or control plasmid. Expression was induced with doxycycline (Dox) for 18 h. Cells were primed with 200 ng/ml LPS for 4h followed by activation with 6.5 µM nigericin, 30 µg/ml imiquimod or 0.025 mg/ml needle toxin for 2 h. Data are represented as mean ± SD, n = 3. b , Western blots of the whole cell lysates from THP-1 NLRP3-KO cells reconstituted with human or zebrafish NLRP3 or zebrafish NLRP3 containing human PYD. Cells were treated with 1 µg/ml LPS for 3 h followed by activation with 20 µM nigericin or 200 µM imiquimod for 1 h. FLAG-tagged NLRP3 constructs and cleaved GSDMD were visualized with corresponding antibodies. c, d, ASC speck formation in HEK293T cells stably expressing YFP-ASC, upon transfection with indicated NLRP3 constructs: human, zebrafish NLRP3 and zebrafish NLRP3 with B30.2-domain deletion (zebrafish NLRP3 ΔB30.2 ) ( c ); wild-type and oligomer-disrupting mutants of zebrafish NLRP3 ( d ). Cells were transfected with the indicated amount of DNA, and the number of ASC specks per image area was calculated 24 h post-transfection. Data are represented as mean ± SD, n = 3.

Article Snippet: Human full-length ASC was cloned into pLenti-EF1a-C-tGFP (Origene, PS100072) and tGFP was exchanged with YFP.

Techniques: Lactate Dehydrogenase Assay, Construct, Control, Plasmid Preparation, Expressing, Activation Assay, Western Blot, Stable Transfection, Transfection

Journal: bioRxiv

Article Title: Structure of zebrafish NLRP3 reveals a novel mode of inflammasome activation

doi: 10.64898/2026.04.17.719140

Figure Lengend Snippet: a , Western blot analysis of NLRP3 expression in reconstituted cell lines in following doxycycline (Dox) induction. Bands corresponding to FLAG-tagged NLRP3 constructs are indicated with arrows. b, c , Western blot analysis of the whole cell lysates from HEK293T cells reconstituted with YFP-ASC and transfected with the indicated NLRP3 constructs, demonstrating relative expression levels of NLRP3 constructs used for ( b ) and ( c ). FLAG-tagged NLRP3 constructs and β-actin were visualized with corresponding antibodies. Cells were transfected with 100 ng DNA per well in a 96-well plate, and the lysates were collected 24 h post-transfection.

Article Snippet: Human full-length ASC was cloned into pLenti-EF1a-C-tGFP (Origene, PS100072) and tGFP was exchanged with YFP.

Techniques: Western Blot, Expressing, Construct, Transfection

Journal: The Biochemical journal

Article Title: FoxA2 and RNA Pol II Mediate Human Islet Amyloid Polypeptide Turnover in ER-stressed Pancreatic β-cells

doi: 10.1042/BCJ20200984

Figure Lengend Snippet: RIN-m5f cells were transduced with empty vector (EV), hIAPP or rIAPP encoding lentivirus particles for 48h. (A) Immuno-confocal microscopy optical section (1μm-Z plane) of RIN-m5f cells transduced with empty vector (EV) and hIAPP encoding lentivirus. Cells were co-stained with hIAPP-specific IAPP monoclonal antibody (red) and DAPI (blue). (B) Representative single (1μm-Z plane) fluorescence confocal sections of hIAPP intracellular accumulation sites in hIAPP lentivirus-transduced cells. Arrows denote hIAPP’s nuclear locations in micrographs. Bars, 5μm.

Article Snippet: Lentiviral particle production and lentivirus-mediated transduction Flag-tagged ORF sequences for pre-pro-hIAPP and rIAPP, flanked with restriction enzyme sites (5’ SgfI and 3’ Mlu1) were synthesized using gBlock gene synthesis (Integrated DNA Technology) and cloned into a third-generation lentivirus destination vector (pLenti-C-Myc-DDK-IRES-Puro, Origene, cat# PS100069).

Techniques: Transduction, Plasmid Preparation, Confocal Microscopy, Staining, Fluorescence