Journal: bioRxiv

Article Title: Inhibition of p107 alleviates liver steatosis by reducing de novo fatty acid synthesis

doi: 10.64898/2026.04.14.718271

Figure Lengend Snippet: (A) p107 protein levels in THLE2 cells transfected with siRNA p107 or siRNA control for 48 hours (n=3 per group). (B) Representative microphotographs of Oil Red O staining (left pannel) of THLE2 cells downregulating p107 (sip107) for 48 hours (n = 3 per group) and Oil Red O semiquantification (right pannel) (n = 3 per group). (C) Quantification of immunoblot analysis of de novo lipogenesis markers (n = 3 per group) and a representative immunoblot. (D) RNA expression of de novo lipogenesis markers (n = 3 per group. (E) De novo synthesis of free fatty acids (FFA) and triglycerides (TG) in THLE2 (n = 6 per group). (F) FAO activity in THLE2 cells transfected with siRNA p107 or siRNA control for 48 hours (n=5 per group). (G) Oxidation rate of palmitic acid (n = 6 per group). (H and I)) OCR and (I) ECAR of p107-silenced THLE2 (n = 17-19 per group). (J) Basal energetic metabolic states, based on quantification of ECAR and OCR during basal metabolism. (K) p107 protein levels in THLE2 after overexpressing p107 (n = 3 per group). (L) Representative microphotographs (left pannel) and semiquantification (right pannel) of Oil Red O staining of THLE2 cells overexpressing p107 (plasmid p107) for 24 hours. Oil Red O staining was quantified using ImageJ and normalized to the total number of nuclei per field (n = 4 per group). (M) Quantification of immunoblot analysis of de novo lipogenesis markers (n = 3 per group) and a representative immunoblot. (N) OCR of p107-overexpressed THLE2 (n =10 per group). GAPDH was used to normalize protein levels. Data are expressed as mean ±SEM. *p < 0.05, **p < 0.01, ***p <0.001, using a Student’s t test .

Article Snippet: THLE2 human hepatic cell line (American Type Culture Collection, ATCC) was cultured in bronchial epithelial cell basal medium (BEBM) supplemented with a growth factors BulleKit (Lonza/Clonetics Corporation), 70ng/mL phosphoethanolamine, 5 ng/mL epidermal growth factor, 10% (v/v) FBS and 1% (v/v) Glutamine-Penicillin-Streptomycin solution (MERCK).

Techniques: Transfection, Control, Staining, Western Blot, RNA Expression, Activity Assay, Plasmid Preparation

Journal: bioRxiv

Article Title: Inhibition of p107 alleviates liver steatosis by reducing de novo fatty acid synthesis

doi: 10.64898/2026.04.14.718271

Figure Lengend Snippet: (A) Volcano plot of total protein expression in p107 liver KO mice compared to shLucif controls (n = 7). Red and blue points indicate significantly up- and downregulated proteins (p < 0.05). (B) Volcano plot of phosphosite abundance in p107 liver KO mice compared to shLucif controls (n = 7). Points represent individual phosphosites—annotated by their parent protein name—with red and blue indicating significant changes (p < 0.05). (C) Heatmap of total protein expression differences grouped by GO Biological Process terms, filtered by significance (p < 0.001). (D) Heatmap of robust total protein expression differences grouped by GO terms, utilizing strict filtering (> 3 combined razor and unique peptides, p < 0.001, absolute t-test difference > 0.58). (E) Quantification of p107 (left panel) and FAS (right panel) in THLE2 after overexpressing p107 (plasmid p107) and silencing FAS (siRNA FAS) during 24h (n = 3 per group) and representative immunoblot. (F) Representative microphotographs of Oil Red O staining of THLE2 cells overexpressing p107 (plasmid p107) and silencing FAS (siRNA FAS) for 24 hours. (G) Semiquantification of Oil Red O staining. Vinculin was used to normalize protein levels. Data are expressed as mean ±SEM. *p < 0.05, **p < 0.01, ***p <0.001, using a Student’s t test .

Article Snippet: THLE2 human hepatic cell line (American Type Culture Collection, ATCC) was cultured in bronchial epithelial cell basal medium (BEBM) supplemented with a growth factors BulleKit (Lonza/Clonetics Corporation), 70ng/mL phosphoethanolamine, 5 ng/mL epidermal growth factor, 10% (v/v) FBS and 1% (v/v) Glutamine-Penicillin-Streptomycin solution (MERCK).

Techniques: Expressing, Phospho-proteomics, Plasmid Preparation, Western Blot, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: Long non-coding RNA H19 transported by colorectal cancer small extracellular vesicles promotes alternative splicing in healthy hepatocytes: new insights on liver pre-metastatic niche formation

doi: 10.1186/s12964-026-02738-x

Figure Lengend Snippet: a qRT-PCR showing lncH19 levels inside CRC-sEVs and CRC_cells. b Representative images of western blot for the presence of RBFOX2 in the CRC-sEVs protein lysates. c Representative Image showing the proposed mechanism of lncH19 acting as shuttle RNA transferring RBFOX2 inside the EVs. d Representative confocal image of RNA in situ assay coupled with immunocytochemistry in the THLE2 treated for 3 h with the CRC-sEVs, lncH19 (red) and RBFOX2 (green). Scale bar:60 μm. The results reported in the graphs are the mean ± SD of three independent experiments. Statistical analyses were performed using Ordinary one-way ANOVA * p < 0.05, ** p < 0.01, *** p < 0.001. e Graphs showing the mean fluorescence intensity (MFI) relative to RBFOX2 (left) and lncH19 (right) of the different conditions

Article Snippet: The SV40 large T antigen-immortalized healthy human liver epithelial cell line THLE2 (ATCC, Manassas, VA) was cultured in Airway Epithelial Cell Basal Medium (ATCC, Manassas, VA) with the Bronchial Epithelial Cell Growth Kit (ATCC PCS-300–040, Manassas, VA) supplemented with 5 ng/ml epidermal growth factor (EGF), 70 ng/ml phosphoethanolamine, 10% fetal bovine serum, 100 U/mL penicillin, and 100 μg/ml streptomycin (Euroclone, UK) at 37 °C with 5% CO 2 .

Techniques: Quantitative RT-PCR, Western Blot, Transferring, In Situ, Immunocytochemistry, Fluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: Long non-coding RNA H19 transported by colorectal cancer small extracellular vesicles promotes alternative splicing in healthy hepatocytes: new insights on liver pre-metastatic niche formation

doi: 10.1186/s12964-026-02738-x

Figure Lengend Snippet: a Representative c onfocal micrographs of RNA in situ hybridisation in THLE2 treated for 18 h with the wt-CRC-sEVs and the sh-CRC-sEVs (lncH19 in red, Hoechst in blue). b MFI analysis obtained with NIS 1 A analysis software, and ( c ) qRT-PCR showing lncH19 levels in treated cells. ( d ) Representative confocal micrographs of immunocytochemistry for RBFOX2 in the THLE2 treated with wt- and sh- sEVs for 18 h and relative Scale bar: 60 μm ( e ) MFI analysis.The results reported in the graphs are the mean ± SD of three independent experiments. Statistical analyses were performed using Ordinary one-way ANOVA *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: The SV40 large T antigen-immortalized healthy human liver epithelial cell line THLE2 (ATCC, Manassas, VA) was cultured in Airway Epithelial Cell Basal Medium (ATCC, Manassas, VA) with the Bronchial Epithelial Cell Growth Kit (ATCC PCS-300–040, Manassas, VA) supplemented with 5 ng/ml epidermal growth factor (EGF), 70 ng/ml phosphoethanolamine, 10% fetal bovine serum, 100 U/mL penicillin, and 100 μg/ml streptomycin (Euroclone, UK) at 37 °C with 5% CO 2 .

Techniques: In Situ, Hybridization, Software, Quantitative RT-PCR, Immunocytochemistry

Journal: Cell Communication and Signaling : CCS

Article Title: Long non-coding RNA H19 transported by colorectal cancer small extracellular vesicles promotes alternative splicing in healthy hepatocytes: new insights on liver pre-metastatic niche formation

doi: 10.1186/s12964-026-02738-x

Figure Lengend Snippet: a qRT-PCR and ( b ) exon-specific qRT-PCR showing the expression levels of ENAH, CTTN and PARD3 genes in THLE2 treated with the wt- and sh-CRC-sEVs

Article Snippet: The SV40 large T antigen-immortalized healthy human liver epithelial cell line THLE2 (ATCC, Manassas, VA) was cultured in Airway Epithelial Cell Basal Medium (ATCC, Manassas, VA) with the Bronchial Epithelial Cell Growth Kit (ATCC PCS-300–040, Manassas, VA) supplemented with 5 ng/ml epidermal growth factor (EGF), 70 ng/ml phosphoethanolamine, 10% fetal bovine serum, 100 U/mL penicillin, and 100 μg/ml streptomycin (Euroclone, UK) at 37 °C with 5% CO 2 .

Techniques: Quantitative RT-PCR, Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: Long non-coding RNA H19 transported by colorectal cancer small extracellular vesicles promotes alternative splicing in healthy hepatocytes: new insights on liver pre-metastatic niche formation

doi: 10.1186/s12964-026-02738-x

Figure Lengend Snippet: a Table showing the different miRNAs identified in the three different analyses (sponged by lncH19, involved in EMT and expressed by healthy liver tissue). b Venn Diagram from FunRich analysis of data obtained by the three indicated datasets. The square in figure indicates 4 from 11 miRNAs targeting PARD3 mRNA c qRT-PCR confirming the over-expression of lncH19 in the THLE2 and ( d ) the increase of PARD3 mRNA in the H19-overexpressing hepatocytes

Article Snippet: The SV40 large T antigen-immortalized healthy human liver epithelial cell line THLE2 (ATCC, Manassas, VA) was cultured in Airway Epithelial Cell Basal Medium (ATCC, Manassas, VA) with the Bronchial Epithelial Cell Growth Kit (ATCC PCS-300–040, Manassas, VA) supplemented with 5 ng/ml epidermal growth factor (EGF), 70 ng/ml phosphoethanolamine, 10% fetal bovine serum, 100 U/mL penicillin, and 100 μg/ml streptomycin (Euroclone, UK) at 37 °C with 5% CO 2 .

Techniques: Quantitative RT-PCR, Over Expression

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