Journal: Cell Death Discovery

Article Title: Detection of inflammasome activation in liver tissue during the donation process as potential biomarker for liver transplantation

doi: 10.1038/s41420-024-02042-y

Figure Lengend Snippet: Representative hematoxylin-eosin images of donated livers biopsies collected during liver procurement depicting subcapsular necrosis, portal space inflammation, and hepatocyte ballooning (black arrows). A control liver with a score of 0 for all analyzed histological parameters is also included.

Article Snippet: White arrowheads indicate the presence of ASC specks. d Liver biopsy samples were double counter-stained with anti-ASC (green) and anti-active IL-1β (red) (Invitrogen; #PA5-105048).

Techniques:

Journal: Cell Death Discovery

Article Title: Detection of inflammasome activation in liver tissue during the donation process as potential biomarker for liver transplantation

doi: 10.1038/s41420-024-02042-y

Figure Lengend Snippet: Representative images of ASC specks, indicating the presence of activated inflammasomes, detected by immunofluorescence in THP-1 cells ( a ) or liver tissue ( b – d ). a THP-1 cells were in vitro primed with LPS for 4 hours and then incubated in the presence or absence of nigericin for an additional 1 hour. Following stimulation, the cells were fixed, centrifuged, and embedded in a specialized processing gel. The gel block was then processed, including fixation in PaxGene medium and paraffin embedding. The white-lined square in the upper right corner represents a magnification of the image. White arrowheads indicate the presence of ASC specks represented as a green dot. The bar-graph depicts the quantification of ASC specks per nucleus in three independent images. *** p ≤ 0.001. b Two different patterns of ASC speck staining can be found in fixed liver tissues. In the upper picture, green dots, indicated by white arrowheads, are detected, while in the lower picture, a reticular structure surrounding the nucleus is shown. A magnification of each image is shown on the right side. Two representative images are shown. c Liver biopsy samples were double counter-stained with anti-ASC antibody (green) and anti-CD68 antibody (red) (Invitrogen; #14-0688-82). White arrowheads indicate the presence of ASC specks. d Liver biopsy samples were double counter-stained with anti-ASC (green) and anti-active IL-1β (red) (Invitrogen; #PA5-105048). Green arrowheads signal single staining for ASC specks, red arrowheads indicate the presence of single staining for active IL-1β, while yellow arrowheads show co-staining for both markers. Two representative images are shown.

Article Snippet: White arrowheads indicate the presence of ASC specks. d Liver biopsy samples were double counter-stained with anti-ASC (green) and anti-active IL-1β (red) (Invitrogen; #PA5-105048).

Techniques: Immunofluorescence, In Vitro, Incubation, Blocking Assay, Staining

Journal: Cell Death Discovery

Article Title: Detection of inflammasome activation in liver tissue during the donation process as potential biomarker for liver transplantation

doi: 10.1038/s41420-024-02042-y

Figure Lengend Snippet: Representative images of the human hepatocyte-like Huh7 cell line transfected with YFP-tagged NLRP3 protein in resting conditions ( a ), stimulated with nigericin ( b ) or incubated with eiOPS from different donated livers (P 1-5 ) or Celsior as negative control ( c ). Representative pictures were taken after 2 hours ( a, b ) or 4 hours (c) of incubation, respectively.

Article Snippet: White arrowheads indicate the presence of ASC specks. d Liver biopsy samples were double counter-stained with anti-ASC (green) and anti-active IL-1β (red) (Invitrogen; #PA5-105048).

Techniques: Transfection, Incubation, Negative Control

Journal: Cell Death Discovery

Article Title: Detection of inflammasome activation in liver tissue during the donation process as potential biomarker for liver transplantation

doi: 10.1038/s41420-024-02042-y

Figure Lengend Snippet: a Representative western blot for the detection of caspase-1 (pro- and active forms) and ASC in liver biopsy extracts from 4 different donated livers collected before donation (T1) or after cold ischemic storage (T2). Β-actin was used as a loading control. Full-length western blots are published as a . b Concentration of IL-1β in the 4 extracted tissues as detected by ELISA. c Correlation matrix among the expression of different NLRP3 inflammasome-related genes detected in liver tissue collected before donation (T1) or after cold ischemic storage (T2).

Article Snippet: White arrowheads indicate the presence of ASC specks. d Liver biopsy samples were double counter-stained with anti-ASC (green) and anti-active IL-1β (red) (Invitrogen; #PA5-105048).

Techniques: Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay, Expressing