Journal: iScience

Article Title: Identification and validation of key PANoptosis-related genes via integrative machine learning and single-cell sequencing in AILI

doi: 10.1016/j.isci.2026.115183

Figure Lengend Snippet: Expression and functional exploration of the key genes in single-cell sequencing data (A) The UMAP plot shows the total sample composition, tissue sources, and cell subtypes. (B) The stacked graph shows the proportion of each type of cell in the control group and the AILI group. (C) Bubble plots of marker gene expression demonstrating the accuracy of the cell annotations. (D) Bubble chart showing the expression of Cdkn1a and Pdk1 in various cells in the control group. (E) Bubble chart showing the expression of Cdkn1a and Pdk1 in various cells in the AILI group. (F) Circle plot and heatmap showing the cell communication weights and numbers of all cell subtypes. (G–J) Receptor‒ligand communication weights between AILI and control samples.

Article Snippet: PDK1 Polyclonal antibody , Proteintech , Cat No.18262-1-AP; RRID: AB_10598310.

Techniques: Expressing, Functional Assay, Single Cell, Sequencing, Control, Marker, Gene Expression

Journal: iScience

Article Title: Identification and validation of key PANoptosis-related genes via integrative machine learning and single-cell sequencing in AILI

doi: 10.1016/j.isci.2026.115183

Figure Lengend Snippet: Validation of key PANoptosis-related genes in animal models (A) H&E staining of liver tissues from WT and AILI mice (scale bars, 100 μm; n = 5). (B) mRNA expression of Cdkn1a and Pdk1 by RT-qPCR. (C–F) Correlation analyses between hepatic Cdkn1a and Pdk1 mRNA levels and serum ALT and AST levels at 24 h after AILI. (G) Detection and statistical analysis of key PANoptosis-related gene and marker protein expression in liver tissues from WT and AILI mice. (H) Immunohistochemical staining for P21 and PDK1 in liver tissues from WT and AILI mice (scale bars, 50 μm; n = 5). All the data are presented as the means ± SDs. One-way ANOVA with Tukey’s test and a two-tailed Student’s t test were used for statistical analysis. Spearman’s rank correlation was used to assess the associations between relative mRNA expression levels of Cdkn1a and Pdk1 and serum ALT and AST levels. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: PDK1 Polyclonal antibody , Proteintech , Cat No.18262-1-AP; RRID: AB_10598310.

Techniques: Biomarker Discovery, Staining, Expressing, Quantitative RT-PCR, Marker, Immunohistochemical staining, Two Tailed Test

Journal: iScience

Article Title: Identification and validation of key PANoptosis-related genes via integrative machine learning and single-cell sequencing in AILI

doi: 10.1016/j.isci.2026.115183

Figure Lengend Snippet: Pdk1 knockdown exacerbates AILI by promoting PANoptosis in vivo Mice were assigned to four groups: control, sh- NC , APAP, and sh- Pdk1 + APAP. (A) Schematic illustration of the experimental design. (B) Western blot analysis confirming efficient knockdown of PDK1 protein in liver tissues. (C) Hematoxylin and eosin (H&E) staining of liver sections and quantification of hepatic necrotic areas. Scale bars, 100 μm; n = 5. (D–E) Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. (F) Serum levels of TNF-α, IL-1β, and IL-6 were measured by ELISA. (G) Western blot analysis and quantification of PANoptosis marker proteins in liver tissues. (H–K) Representative immunofluorescence staining of liver sections showing albumin (ALB, green) and PANoptosis marker proteins (ZBP1, p -MLKL, cleaved caspase-1, and cleaved caspase-3; red). Scale bars, 20 μm; n = 5. Data are presented as mean ± SD. One-way ANOVA with Tukey’s test and a two-tailed Student’s t test were used for statistical analysis. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: PDK1 Polyclonal antibody , Proteintech , Cat No.18262-1-AP; RRID: AB_10598310.

Techniques: Knockdown, In Vivo, Control, Western Blot, Staining, Enzyme-linked Immunosorbent Assay, Marker, Immunofluorescence, Two Tailed Test

Journal: bioRxiv

Article Title: Ubiquitin ligase CHFR impairs Tie2 signaling via K 48 -linked ubiquitylation and degradation of Akt1 in endothelial cells

doi: 10.64898/2026.03.31.715582

Figure Lengend Snippet: a-b CHFR deficiency in EC prevents LPS-induced ubiquitylation of Akt1. HLMVEC transfected with sc-siRNA or CHFR-siRNA. At 72 h post-transfection, cells were challenged with LPS (5 μg/ml) for 6 h in the presence of the proteasomal inhibitor MG132 (10 μM). Cell lysates were immunoprecipitated with anti-Akt1 mAb and blotted with antibodies specific to K 48 -linked poly-Ub or K 63 -linked poly-Ub ( n = 2 independent experiments) ( a ). Chfr fl/fl (WT) and Chfr ΔEC mice were challenged with LPS (10 mg/kg; i.p.) for 6 h. After the LPS challenge, lungs harvested were used to determine ubiquitylation of Akt1 as above in a (b) . c-f CHFR induces ubiquitylation of activated Akt1. c Control and CHFR-depleted HLMVEC were pretreated with inhibitors of PDK1 and mTORC2 for 30 min and then exposed to LPS (5 μg/ml). Thereafter, cell lysates were used for IB to assess phosphorylation of Akt1. d-e HLMVEC pretreated with or without PDK1 and mTORC2 inhibitors for 1 h and stimulated with LPS (6 h) in the presence of MG123 (10 μM) showed that CHFR binds and ubiquitylates phosphorylated Akt1. f HEK-293T cells were transfected with HA-tagged ubiquitin (HA-Ub) (0.5 μg/ml) alone or co-transfected with N-terminal GFP-tagged WT-CHFR (1.5 μg/ml), N-terminal pmCherry-tagged WT-Akt1 (1.5 μg/ml), and phosphorylation-defective Akt1 mutant (Akt1T308A/S473A) (1.5 μg/ml) plasmids. Thirty-six hours after transfection, cells were incubated with MG123 (10 μM) for 3h, and cell lysates were used to determine phosphorylation-dependent ubiquitylation of Akt1.

Article Snippet: Mammalian targets of rapamycin (mTOR) inhibitor (catalog #WYE-354), and 3-Phosphoinositide-Dependent Kinase 1 (PDK1) inhibitor (catalog #GSK2334470) were from Selleckchem.com.

Techniques: Transfection, Immunoprecipitation, Control, Phospho-proteomics, Ubiquitin Proteomics, Mutagenesis, Incubation