human adipose derived mesenchymal stem cells asc  (ATCC)

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 , Position and sequence of the PYD-FISNA linker in human and zebrafish NLRP3, and zebrafish NLRP3 containing human PYD-FISNA linker sequence encoded by the exon 3 of human NLRP3 gene (red dashed box). Sequences corresponding to FISNA domain and polybasic region are indicated with black lines. Cys130 palmitoylated in human NLRP3 is highlighted in blue. b , SDS-PAGE gels of sucrose gradient fractions of zebrafish NLRP3 WT and zebrafish NLRP3 containing human linker sequence. c , Sequence alignment of zebrafish and human NLRP3. Domains are indicated with lines and color-coded. Residues forming a “face-to-face” interface according to the human (PDB: 7PZC) and mouse (PDB: 7LFH) NLRP3 “cage” structures are indicated with red and orange stars, respectively. Residues forming a “back-to-back” interface according to the human (PDB: 7PZC) and mouse (PDB: 7LFH) NLRP3 “cage” structures are indicated with blue and violet stars, respectively. Numbers correspond to alignment positions within the full-length sequence alignment. d , Position and sequence of known palmitoylation sites in human and zebrafish NLRP3. Palmitoylated residues are highlighted in blue.

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

Techniques: Sequencing, SDS Page

Journal: PLOS Pathogens

Article Title: The non-structural protein of SFTSV activates NLRP1 and CARD8 inflammasome through disrupting the DPP9-mediated ternary complex

doi: 10.1371/journal.ppat.1013258

Figure Lengend Snippet: (A-B) ASC specks fluorescence microscopy images ( A ) and quantification ( B ) of A549-HA-NLRP1-Flag-ASC-GFP cells infected with SFTSV or without SFTSV (MOI = 0.5) for 24 h. Scale bar, 100 μm. (C-D) Detection of SFTSV NP ( C ) and IL-β production ( D ) in primary keratinocytes infected with SFTSV at different MOIs or stimulated with VbP (2 μM) for 24 h. (E-F) GSDMD cleavage ( E ) and LDH release ( F ) in primary keratinocytes infected with SFTSV (MOI = 1) or stimulated with VbP (5 μM) for 36 h. (G) Expression of NLRP1 in primary keratinocytes treated with lentivirus-mediated CRISPR-Cas9 and NLRP1 -specific single-guide RNA (sgRNA) or the non-targeting sgRNA control. (H) IL-1β production in wild-type or NLRP1-deficient primary keratinocytes infected with SFTSV (MOI = 1) or stimulated with VbP (2 μM) for 24 h. (I) Primary keratinocytes were infected with SFTSV at different MOIs for 24 h, endogenous NLRP1 was detected using N-terminal and C-terminal antibodies with Western blot, respectively. (J-K) Primary keratinocytes were infected with SFTSV (MOI = 1) and treated with MG132 (2 μM) for 24 h, IL-1β release ( J ) in the cell supernatant was measured with ELISA; endogenous NLRP1 ( K ) was detected with Western blot. All data represent three independent experiments and presented as mean±s.d. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns , not significant. For statistical analysis, one-way ANOVA in (D, J), two-tailed unpaired Student’s t -test in (B, F, H).

Article Snippet: Human NLRP1 (HG30111-NY), ASC (HG11175-CY), pro-Casp-1 (HG11148-NF), GSDMD (HG25207-NF), pro-IL-1β (HG10139-NM), CARD8 (HG12619-NM) and DPP9 (HG11418-NF) plasmid and the ASC-GFP (HG11175-ACGLN) lentiviral vector were purchased from Sino Biological (Beijing, China).

Techniques: Fluorescence, Microscopy, Infection, Expressing, CRISPR, Control, Western Blot, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: PLOS Pathogens

Article Title: The non-structural protein of SFTSV activates NLRP1 and CARD8 inflammasome through disrupting the DPP9-mediated ternary complex

doi: 10.1371/journal.ppat.1013258

Figure Lengend Snippet: (A) ASC-caspase-1-pro-IL-1β HEK293T cells were transfected with indicated expression vector for 36 h. Supernatants were analyzed for IL-1β with ELISA. (B) ASC-caspase-1-pro-IL-1β HEK293T cells were transfected with plasmids NSs, production of p17 and GSDMD processing were detected with Western blot. (C-D) ASC specks fluorescence microscopy images ( C ) and quantification ( D ) of A549-HA-NLRP1-Flag-ASC-GFP cells transfected NSs-HA for 24 h. Scale bar, 100 μm. (E-F) GSDMD cleavage ( E ) and IL-1β ( F ) production in primary keratinocytes expressing Tet-HA-NSs and treated with doxycycline (1 μg/ml) for 48 h. (G-H) GSDMD cleavage ( G ) and IL-1β ( H ) production in THP-1 cells expressing Tet-HA-NSs and treated with doxycycline (1 μg/ml) for 48 h. All data represent three independent experiments and presented as mean±s.d. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns , not significant. For statistical analysis, two-tailed unpaired Student’s t -test in (A, D, F, H).

Article Snippet: Human NLRP1 (HG30111-NY), ASC (HG11175-CY), pro-Casp-1 (HG11148-NF), GSDMD (HG25207-NF), pro-IL-1β (HG10139-NM), CARD8 (HG12619-NM) and DPP9 (HG11418-NF) plasmid and the ASC-GFP (HG11175-ACGLN) lentiviral vector were purchased from Sino Biological (Beijing, China).

Techniques: Transfection, Expressing, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Western Blot, Fluorescence, Microscopy, Two Tailed Test