Journal: iScience

Article Title: FOXE1 promotes the progression of pulp inflammation by activating PANoptosis in dental pulp cells

doi: 10.1016/j.isci.2026.115204

Figure Lengend Snippet: Mixed infection with S.m. and F.n. induced PANoptosis in DPCs (A–H) Dental pulp cells were infected with S.m. (MOI = 100), F.n. (MOI = 100), or a combination of S.m. (MOI = 100) and F.n. (MOI = 100) or treated with PBS (negative control, NC) for 8 h. (A) Dental pulp cells were treated with PBS, S.m. , F.n. , or a combination of S.m. and F.n. ( n = 5 experiments in each group), and cell death was quantified by measuring LDH release. (B) Dental pulp cells were treated with PBS, S.m. , F.n. , or a combination of S.m. and F.n . Treated DPCs were stained with PI, fixed, counterstained with DAPI, and visualized under a microscope. Scale bars, 100 μm. (C) Quantification of the proportions of PI-positive cells in B ( n = 6 experiments in each group). (D) Dental pulp cells were treated as indicated and analyzed by flow cytometry. (E) PANoptotic DPCs were quantified by determining the percentages of PI + /annexin V + cells ( n = 3 experiments in each group). (F) The protein levels of cleaved caspase-3 p17, cleaved GSDMD NT, and p -MLKL were assessed by Western blot. (G) Immunofluorescence staining of PANoptotic markers in infected DPCs, as visualized using a confocal microscope. Scale bars, 10 μm. (H) Quantification of PANoptotic cell proportions in DPCs treated with PBS, S.m. , F.n. , or a combination of S.m. and F.n. ( n = 8 samples in each group), corresponding to (G). Data are represented as mean ± SEM. ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 by one-way ANOVA (A, C, and E) or by Kruskal-Wallis test (H).

Article Snippet: The following primary antibodies were used in this study: human and mouse vimentin (1:200, #ab92547, Abcam), human cleaved caspase-3 (1:100, #9664, CST), human cleaved GSDMD (1:100, #36425, CST), human p-MLKL (1:100, #PA5-105678, Invitrogen), mouse cleaved caspase-3 (1:100, #BF0711, Affinity), mouse cleaved GSDMD (1:100, #ab255603, Abcam), mouse p-MLKL (1:100, #ab196436, Abcam) and mouse FOXE1 (1:50, #55363-1-AP, Proteintech).

Techniques: Infection, Negative Control, Staining, Microscopy, Flow Cytometry, Western Blot, Immunofluorescence

Journal: iScience

Article Title: FOXE1 promotes the progression of pulp inflammation by activating PANoptosis in dental pulp cells

doi: 10.1016/j.isci.2026.115204

Figure Lengend Snippet: Identification and validation of FOXE1 as a key transcription factor that regulates PANoptosis in DPCs (A) Differentially expressed gene analysis was performed between the PANoptotic DPC cluster (cluster 1) and other DPC clusters (clusters 0, 2, 4, 5, and 6) and between cluster 1 and all remaining clusters. The candidate regulators were selected from the overlapping DEGs between these comparisons. (B) The top five candidate regulators were identified after comparison. (C) siRNA targeting FOXE1 (20 μM) was transfected into DPCs with Lipofectamine 2000 for 24 h. The knockdown efficiency was validated by qRT-PCR ( n = 3 experiments in each group) and Western blot. (D–H) Dental pulp cells were pretreated with siRNAs (20 μM) targeting candidate regulators for 24 h, followed by infection with S.m. (MOI = 100) and F.n. (MOI = 100) for 8 h. (D) Cell death was quantified by the LDH release assays ( n = 5 experiments in each group). (E, also see in ) Cell death was assessed by PI staining ( n = 4 experiments in each group). (F) Treated cells were analyzed by flow cytometry. (G) PANoptotic DPCs were quantified by determining the percentages of PI + /annexin V + cells ( n = 3 experiments in each group). (H) Western blot analysis was used to assess the expression of cleaved caspase-3 p17, cleaved GSDMD NT and p -MLKL. (I) Immunofluorescence staining of PANoptotic markers in si FOXE1 -and siCON-transfected DPCs. Scale bars, 10 μm. (J) Quantification of PANoptotic DPCs in I ( n = 5 samples in each group). Data are represented as mean ± SEM. ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 by Student’s t test (C, G, and J) or by one-way ANOVA (D and E).

Article Snippet: The following primary antibodies were used in this study: human and mouse vimentin (1:200, #ab92547, Abcam), human cleaved caspase-3 (1:100, #9664, CST), human cleaved GSDMD (1:100, #36425, CST), human p-MLKL (1:100, #PA5-105678, Invitrogen), mouse cleaved caspase-3 (1:100, #BF0711, Affinity), mouse cleaved GSDMD (1:100, #ab255603, Abcam), mouse p-MLKL (1:100, #ab196436, Abcam) and mouse FOXE1 (1:50, #55363-1-AP, Proteintech).

Techniques: Biomarker Discovery, Comparison, Transfection, Knockdown, Quantitative RT-PCR, Western Blot, Infection, Staining, Flow Cytometry, Expressing, Immunofluorescence

Journal: iScience

Article Title: CGF induces ROS-mediated metabolic reprogramming and mitochondrial dysfunction to suppress colorectal cancer progression

doi: 10.1016/j.isci.2026.115273

Figure Lengend Snippet: CGF’s effect on cell cycle and apoptosis in CRC (A) Flow cytometry was used to analyze how CGF affects the cell cycle of HCT116 and HT29 cells at certain concentrations, with the percentage of cells in G1, S, and G2 phases shown in each panel. (B) Western blot analysis of the changes in cell cycle-related proteins CDK1, p-CDK1, and cyclin B1 in HCT116 and HT29 cells after CGF treatment. (C) RT-qPCR analysis of the relative expression levels of PUMA and NOXA genes in HCT116 and HT29 cells treated with different concentrations of CGF. (D) Western blot analysis of the changes in apoptosis-related proteins BCL2, PUMA, Noxa, C-caspase 9, and C-caspase 3 in HCT116 and HT29 cells after CGF treatment. (E) Flow cytometry was used to analyze apoptosis in HCT116 and HT29 cells treated with CGF. On the left is a representative plot showing apoptosis, utilizing Annexin V-FITC and PI double staining. Right: Analysis of early and late apoptosis in cells from each group using quantitative methods. (A–C and E) Data presentation is in the form of mean ± SEM. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb , CST , Cat # 9664; RRID:AB_2070042.

Techniques: Flow Cytometry, Western Blot, Quantitative RT-PCR, Expressing, Double Staining

Journal: iScience

Article Title: CGF induces ROS-mediated metabolic reprogramming and mitochondrial dysfunction to suppress colorectal cancer progression

doi: 10.1016/j.isci.2026.115273

Figure Lengend Snippet: The effect of CGF-induced ROS on the MAPK/ERK 1/2/c-MYC signaling pathway (A) Western blotting was used to analyze the expression of MEK, P-MEK, ERK 1/2, P-ERK 1/2, and c-MYC proteins in HCT116 and HT29 cells after treatment with specified CGF concentrations. (B) Immunofluorescence staining of HCT116 and HT29 cells after CGF treatment, showing changes in p-ERK (green). PMA was used to promote nuclear entry of p-ERK (200 nM, 4 h), and DAPI was used to stain the nucleus (blue), with merged images shown (scale bars, 10 μM). (C) Western blot was used to analyze the levels of MEK, P-MEK, ERK 1/2, P-ERK 1/2, and c-MYC proteins in HCT116 and HT29 cells following treatment with CGF (40 μM, 24 h) and NAC (500 μM, 24 h). (D) Untargeted metabolomics analysis of glycolytic metabolites after CGF treatment in HCT116 cells. Metabolites marked in blue represent those inhibited by CGF. (E and F) ELISA analysis of lactate secretion in the cell supernatant of HCT116 (E) and HT29 (F) cells after CGF treatment. (G and H) Analysis of mRNA expression levels of genes related to glycolysis in HCT116 and HT29 cells exposed to certain concentrations of CGF using RT-qPCR. (I) Western blot was used to analyze the expression levels of apoptosis-associated proteins (BCL2, PUMA, NOXA, C-caspase 9, and C-caspase 3) and EMT markers (N-cadherin and vimentin) in HCT116 and HT29 cells after they were treated with CGF. (J) Western blot analysis of the reversal of EMT markers N-cadherin, vimentin, SLUG, and SNAIL proteins in HCT116 and HT29 cells overexpressing c-MYC. (E–J) Data presentation is in the form of mean ± SEM. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb , CST , Cat # 9664; RRID:AB_2070042.

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR