human immortalized hepatocytes thle2  (ATCC)

Journal: Genome Biology

Article Title: O-GlcNAcylation of NONO mediates alternative splicing of SETMAR and facilitates NHEJ repair

doi: 10.1186/s13059-026-03930-5

Figure Lengend Snippet: O-GlcNAcylation of NONO promotes radioresistance in hepatocellular carcinoma. A Analysis of NONO O-GlcNAcylation was conducted in human liver epithelial cells (THLE2) and hepatocellular carcinoma cells (HepG2, HCCLM9, Huh7) usingIP/Western blotting with indicated antibodies. B Generation of stable HCCLM9 cells with NONO knockdown. Western blot analysis was performed to assess the efficiency of NONO knockdown using two shRNA constructs(shNONO #1 and shNONO #2). GAPDH was used as a loading control. C Upper panel: O-GlcNAcylation at Ser147 promotes cell survival post-IR treatment. NONO-knockdown HCCLM9 cells were transfected with SFB-NONO WT or S147A mutant and subjected to clonogenic survival assays post-IR treatment. Cells were treated with the indicated doses of IR and further incubated for 7–10 days. Lower panel: Quantitative analysis of clonogenic survival assays. D Schematic diagram illustrating the radiotherapy process for NTG mice. Mice injected with control or NONO knockdown cells, as well as cells reconstituted with NONO WT or S147A and wereexposed to 8 Gy of IR twice. E O-GlcNAcylation of NONO enhances radioresistance. Mice were subcutaneously injected with 7 × 10 6 control or NONO knockdown cells, or cells reconstituted with NONO WT or S147A, and exposed to 8 Gy of IR twice or not, when tumors reached a similar size (about 100 mm. 3 ). Representative images of xenograft tumors are shown ( n = 6/group). F Quantification analysis of xenograft tumor volumes from ( E ). Data are presented as mean ± SD. Statistical significance was determined using one-way ANOVA. ** P < 0.01, *** P < 0.001. G Hematoxylin and eosin (H&E) staining and IHC analysis of NONO and H3K36me2 in xenograft tumors were performed, comparing controland NONO knockdown groups, as well as tumors reconstituted with NONO WT or S147A ( n = 6). Scale bar, 30 μm. H Quantification of H3K36me2 levels from ( G ). Data are presented as mean ± SD. Statistical significance was determined using one-way ANOVA. ** P < 0.01, *** P < 0.001

Article Snippet: The HEK293T, U2OS, THLE2, Huh7, HepG2 and HCCLM9 cell lines were purchased from American Type Cell Culture (ATCC), and cultured in Dulbecco’s modified Eagle medium (Gibco) supplemented with 10% fetal bovine serum (FBS) (Gibco), 100 U/mL penicillin, and 100 μg/mL streptomycin in the presence of 5% CO 2 (v/v) at 37 °C.

Techniques: Western Blot, Knockdown, shRNA, Construct, Control, Transfection, Mutagenesis, Incubation, Injection, Staining

Journal: Frontiers in Pharmacology

Article Title: Ultrafine garlic powder alleviates non-alcoholic steatohepatitis by inhibiting hepatocyte ferroptosis and modulating ERK-dependent oxidative stress

doi: 10.3389/fphar.2025.1711917

Figure Lengend Snippet: UGP ameliorates palmitatic acid-induced lipotoxic hepatocyte injury. Effects of palmitatic acid stimulation and Fer-1, UGP and TGP treatment of on human normal hepatocyte THLE2 cell viability was examined using CCK8 assay (A) and LDH assay (B) (n = 4). (C) Effects of palmitatic acid stimulation and Fer-1, UGP and TGP treatment of on THLE2 cell damage was determined by ALT assay (n = 4). (D) Cell death assay was performed using Annexin V/PI staining (n = 4). Annexin V + PI − , Annexin V + PI − , and Annexin V + PI + cells were considered dead cells. Representative fluorescent images and quantification of DCFDA (2′,7′-dichlorofluorescein diacetate) (E) and Mito-Tracker Red CMXRos (F) staining for ROS and mitochondrial membrane potential detection (scale bar = 100 μm, n = 4). (G) Flow cytometry analysis of lipid peroxidation by Liperfluo staining on THLE2 cells. (H) Representative immunofluorescent images of ACTA2 +ve and COL1A1 +ve cells (scale bar = 100 μm). Data are shown as box-and whisker with median (middle line), 25th–75th percentiles (box), and min-max values (whiskers), one-way ANOVA with Tukey’s correction. (ALT: Alanine transaminase; BSA:Bovine serum albumin; Fer-1: Ferrostatin-1; PAL: Palmitic acid; ROS: Reactive oxygen species; TGP: Traditional garlic powder; UGP:Ultrafine garlic powder).

Article Snippet: Human normal hepatocytes (THLE2) and hepatic stellate cells (HSCs) (LX-2) were purchased from the American Type Culture Collection (ATCC) and cultured in BEGM kit medium and RPMI medium supplemented with 10% FBS, respectively.

Techniques: CCK-8 Assay, Lactate Dehydrogenase Assay, Staining, Membrane, Flow Cytometry, Whisker Assay

Journal: Frontiers in Pharmacology

Article Title: Ultrafine garlic powder alleviates non-alcoholic steatohepatitis by inhibiting hepatocyte ferroptosis and modulating ERK-dependent oxidative stress

doi: 10.3389/fphar.2025.1711917

Figure Lengend Snippet: UGP significantly ameliorates erastin-induced hepatocyte ferroptosis. Effects of erastin stimulation and Fer-1, UGP and TGP treatment of on THLE2 cell viability was examined using CCK8 assay (A) and LDH assay (B) (n = 4). (C) Effects of erastin stimulation and Fer-1, UGP and TGP treatment of on THLE2 cell damage was determined by ALT assay (n = 4). (D) Cell death assay was performed using Annexin V/PI staining (n = 4). Annexin V + PI − , Annexin V + PI − , and Annexin V + PI + cells were considered dead cells. (E) Flow cytometry analysis of cellular Fe 2+ levels by FerroOrange staining on THLE2 cells. Representative fluorescent images and quantification of DCFDA (2′,7′-dichlorofluorescein diacetate) (F) and Mito-Tracker Red CMXRos (G) staining for ROS and mitochondrial membrane potential detection (scale bar = 100 μm, n = 4). (H) Flow cytometry analysis of lipid peroxidation by Liperfluo staining on THLE2 cells. Data are shown as box-and whisker with median (middle line), 25th-75th percentiles (box), and min-max values (whiskers), one-way ANOVA with Tukey’s correction. (ALT: Alanine transaminase; ERA: Erastin; Fer-1: Ferrostatin-1; ROS: Reactive oxygen species; TGP: Traditional garlic powder; UGP: Ultrafine garlic powder).

Article Snippet: Human normal hepatocytes (THLE2) and hepatic stellate cells (HSCs) (LX-2) were purchased from the American Type Culture Collection (ATCC) and cultured in BEGM kit medium and RPMI medium supplemented with 10% FBS, respectively.

Techniques: CCK-8 Assay, Lactate Dehydrogenase Assay, Staining, Flow Cytometry, Membrane, Whisker Assay