Journal: Experimental and Therapeutic Medicine

Article Title: PDK1/AKT signaling is involved in the anti-tumor effect of gramine and cisplatin chemoresistance in ovarian cancer cells

doi: 10.3892/etm.2026.13273

Figure Lengend Snippet: PDK1 overexpression impaired the anti-proliferation effect of gramine on OC cells. (A) Western blotting assay determining the effect of gramine on PDK1 expressions. SK-OV-3 and OV-90 cells were incubated with gramine (50, 100 µM) for 24 h. * P<0.05, ** P<0.01, *** P<0.001 vs. the untreated control. (B) Western blotting assay assessing the transfection efficiency of PDK1 overexpression lentivirus. *** P<0.001 vs. the EV control. (C) Cell counting kit-8 assay detecting the effect of PDK1 overexpression on cell viability. (D) Clone formation assay assessing the effect of PDK1 overexpression on colony formation ability. (E) Western blotting assay examining the effect of PDK1 overexpression on Ki67 expression. * P<0.05, ** P<0.01 vs. the EV control or gramine-treated EV group (C-E). Data represent the mean ± SD of three independent experiments. EV, empty vector; OV, overexpression.

Article Snippet: The primary antibodies used were as follows: Ki67 (1:2,000; 28074-1-AP; Proteintech Group, Inc.), Bax (1:20,000; 50599-2-Ig; Proteintech Group, Inc.), Bcl-2 (1:5,000; 68103-1-Ig; Proteintech Group, Inc.), PARP1 (1:1,000; 13371-1-AP; Proteintech Group, Inc.), cleaved PAPR1 (1:5,000; 60555-1-Ig; Proteintech Group, Inc.), N-cadherin (1:20,000; 22018-1-AP; Proteintech Group, Inc.), E-cadherin (1:20,000; 20874-1-AP; Proteintech Group, Inc.), vimentin (1:20,000; 10366-1-AP; Proteintech Group, Inc.), GAPDH (1:50,000; 60004-1-Ig; Proteintech Group, Inc.), PDK1 (1:1,000; A0834; Abclonal Biotech Co., Ltd.), AKT (1:1,000; 4691; Cell Signaling Technology, Inc.) and p-AKT (Thr308) (1:1,000; 9275; Cell Signaling Technology, Inc.).

Techniques: Over Expression, Western Blot, Incubation, Control, Transfection, Cell Counting, Tube Formation Assay, Colony Assay, Expressing, Plasmid Preparation

Journal: Experimental and Therapeutic Medicine

Article Title: PDK1/AKT signaling is involved in the anti-tumor effect of gramine and cisplatin chemoresistance in ovarian cancer cells

doi: 10.3892/etm.2026.13273

Figure Lengend Snippet: PDK1 overexpression impaired the pro-apoptotic effect of gramine on ovarian cancer cells. (A) TUNEL assay evaluating the effect of PDK1 overexpression on cell apoptosis. Scale bar, 200 µm. (B) Western blotting assay determining the effect of PDK1 overexpression on Bax and Bcl2 expressions. PDK1 overexpressing SK-OV-3 and OV-90 cells were treated with gramine (100 µM) for 24 h. * P<0.05, ** P<0.01, *** P<0.001 vs. the EV control or gramine-treated EV group. Data represent the mean ± SD of three independent experiments. EV, empty vector; OV, overexpression.

Article Snippet: The primary antibodies used were as follows: Ki67 (1:2,000; 28074-1-AP; Proteintech Group, Inc.), Bax (1:20,000; 50599-2-Ig; Proteintech Group, Inc.), Bcl-2 (1:5,000; 68103-1-Ig; Proteintech Group, Inc.), PARP1 (1:1,000; 13371-1-AP; Proteintech Group, Inc.), cleaved PAPR1 (1:5,000; 60555-1-Ig; Proteintech Group, Inc.), N-cadherin (1:20,000; 22018-1-AP; Proteintech Group, Inc.), E-cadherin (1:20,000; 20874-1-AP; Proteintech Group, Inc.), vimentin (1:20,000; 10366-1-AP; Proteintech Group, Inc.), GAPDH (1:50,000; 60004-1-Ig; Proteintech Group, Inc.), PDK1 (1:1,000; A0834; Abclonal Biotech Co., Ltd.), AKT (1:1,000; 4691; Cell Signaling Technology, Inc.) and p-AKT (Thr308) (1:1,000; 9275; Cell Signaling Technology, Inc.).

Techniques: Over Expression, TUNEL Assay, Western Blot, Control, Plasmid Preparation

Journal: Experimental and Therapeutic Medicine

Article Title: PDK1/AKT signaling is involved in the anti-tumor effect of gramine and cisplatin chemoresistance in ovarian cancer cells

doi: 10.3892/etm.2026.13273

Figure Lengend Snippet: PDK1/AKT signaling pathway was involved in the anti-migratory and anti-invasive effects of gramine in ovarian cancer cells. (A) Wound healing and (B) transwell invasion assays detecting the effect of PDK1 overexpression on the abilities of cell migration and invasion respectively. Scale bar, 200 µm. (C) Western blotting assay determining the effect of PDK1 overexpression on N-cadherin, vimentin and p-AKT (Thr308) expressions. PDK1 overexpressing SK-OV-3 and OV-90 cells were incubated with gramine (50 µM) for 24 h. * P<0.05, ** P<0.01, *** P<0.001 vs. the EV control or gramine-treated EV group. Data represent the mean ± SD of three independent experiments. EV, empty vector; OV, overexpression.

Article Snippet: The primary antibodies used were as follows: Ki67 (1:2,000; 28074-1-AP; Proteintech Group, Inc.), Bax (1:20,000; 50599-2-Ig; Proteintech Group, Inc.), Bcl-2 (1:5,000; 68103-1-Ig; Proteintech Group, Inc.), PARP1 (1:1,000; 13371-1-AP; Proteintech Group, Inc.), cleaved PAPR1 (1:5,000; 60555-1-Ig; Proteintech Group, Inc.), N-cadherin (1:20,000; 22018-1-AP; Proteintech Group, Inc.), E-cadherin (1:20,000; 20874-1-AP; Proteintech Group, Inc.), vimentin (1:20,000; 10366-1-AP; Proteintech Group, Inc.), GAPDH (1:50,000; 60004-1-Ig; Proteintech Group, Inc.), PDK1 (1:1,000; A0834; Abclonal Biotech Co., Ltd.), AKT (1:1,000; 4691; Cell Signaling Technology, Inc.) and p-AKT (Thr308) (1:1,000; 9275; Cell Signaling Technology, Inc.).

Techniques: Over Expression, Migration, Western Blot, Incubation, Control, Plasmid Preparation

Journal: Experimental and Therapeutic Medicine

Article Title: PDK1/AKT signaling is involved in the anti-tumor effect of gramine and cisplatin chemoresistance in ovarian cancer cells

doi: 10.3892/etm.2026.13273

Figure Lengend Snippet: Gramine downregulates PDK1/AKT linked to cisplatin sensitization of OC cells. (A) CCK-8 assay detecting the effect of cisplatin on cell viability. SK-OV-3 and SK-OV-3-R cells were incubated with cisplatin (5, 10, 20, 40, 80 µM) for 24 h. * P<0.05, ** P<0.01 vs. the untreated control. (B) CCK-8 assay detecting the effect of gramine and cisplatin on cell viability. SK-OV-3-R cells were incubated with gramine (50 µM) and cisplatin (5, 10, 20, 40, 80 µM) for 24 h. * P<0.05 vs. the DMSO group at the identical cisplatin concentration. (C) Clone formation assay assessing the effect of gramine and cisplatin on colony formation ability. (D) TUNEL assay evaluating the effect of gramine and cisplatin on cell apoptosis. Scale bar, 200 µm. (E) Wound healing and transwell invasion assays detecting the effect of gramine and cisplatin on the abilities of cell migration and invasion respectively. Scale bar, 200 µm. (F) Western blotting assay determining the effect of gramine and cisplatin on the expressions of PDK1 and p-AKT (Thr308). SK-OV-3-R cells were incubated with gramine (50 µM) and cisplatin (80 µM) for 24 h. * P<0.05, ** P<0.01, *** P<0.001 vs. the cisplatin group (C-F). Data represent the mean ± SD of three independent experiments. CCK-8, cell counting kit-8.

Article Snippet: The primary antibodies used were as follows: Ki67 (1:2,000; 28074-1-AP; Proteintech Group, Inc.), Bax (1:20,000; 50599-2-Ig; Proteintech Group, Inc.), Bcl-2 (1:5,000; 68103-1-Ig; Proteintech Group, Inc.), PARP1 (1:1,000; 13371-1-AP; Proteintech Group, Inc.), cleaved PAPR1 (1:5,000; 60555-1-Ig; Proteintech Group, Inc.), N-cadherin (1:20,000; 22018-1-AP; Proteintech Group, Inc.), E-cadherin (1:20,000; 20874-1-AP; Proteintech Group, Inc.), vimentin (1:20,000; 10366-1-AP; Proteintech Group, Inc.), GAPDH (1:50,000; 60004-1-Ig; Proteintech Group, Inc.), PDK1 (1:1,000; A0834; Abclonal Biotech Co., Ltd.), AKT (1:1,000; 4691; Cell Signaling Technology, Inc.) and p-AKT (Thr308) (1:1,000; 9275; Cell Signaling Technology, Inc.).

Techniques: CCK-8 Assay, Incubation, Control, Concentration Assay, Tube Formation Assay, Colony Assay, TUNEL Assay, Migration, Western Blot, Cell Counting

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