normal hepatocyte cell line thle 2  (ATCC)

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: Communications Biology

Article Title: CRISPR-Cas9 screening identifies ATOX1-driven cisplatin resistance mechanisms in liver cancer and evaluates targeted inhibitor efficacy

doi: 10.1038/s42003-026-09722-8

Figure Lengend Snippet: A Schematic diagram of CRISPR-Cas9 gene screen model in HepG2 and Huh7 liver cancer cell lines. B , C sgRNA number distribution of HepG2 and Huh7 cells on day 0. D KEGG pathway analysis of cisplatin resistance genes in HepG2 cells. E KEGG pathway analysis of cisplatin resistance genes in Huh7 cells. F 67 cisplatin resistance genes shared common between HepG2 and Huh7 cells. G Z-score plots indicated the cisplatin resistance genes with top10 drugZ score in two liver cancer cell lines. H Changes of targeting ATOX1 sgRNAs in two liver cancer cell lines in day 0, DMSO, and cisplatin treatment group. I Expression of ATOX1 protein was detected in human immortalized hepatocyte cell line THLE-2 and various liver cancer cell lines, including HepG2, Huh1, Hep3B, SNU449, JHH7, Huh7, Li-7, and SNU-475 ( n = 3). J CCK-8 was used to detect the IC50 of cisplatin in liver cancer cell lines ( n = 3). K Liver cancer cell lines with high ATOX1 protein expression exhibited a high IC50 of cisplatin. ** P < 0.01.

Article Snippet: The human immortalized hepatocyte cell line THLE-2, along with the liver cancer cell lines Huh1, Hep3B, JHH7, and Li-7, were sourced from Procell (Wuhan, China).

Techniques: CRISPR, Expressing, CCK-8 Assay

Journal: Communications Biology

Article Title: CRISPR-Cas9 screening identifies ATOX1-driven cisplatin resistance mechanisms in liver cancer and evaluates targeted inhibitor efficacy

doi: 10.1038/s42003-026-09722-8

Figure Lengend Snippet: A Cell survival curves and IC50 values of compound 8 and DC_AC50 on THLE-2, HepG2 and Huh7 when used alone ( n = 3). B , C Heat map demonstrated the synergistic effects at different concentration combinations ( n = 3). D EDU positive rate in DMSO, compound 8, cisplatin and combine treatment group ( n = 3). White bar means 100 μm. E Colony formation assay showing clonogenic ability in DMSO, compound 8, cisplatin and combine treatment group ( n = 3). F Annexin V/PI double staining flow cytometry analysis of apoptosis rates in DMSO, compound 8, cisplatin and combine treatment group ( n = 3). G Research design of HepG2 liver cancer cell xenograft model. H Tumor volume growth curves of DMSO, compound 8, cisplatin and combine treatment group ( n = 5/group). I In vivo imaging results of DMSO, compound 8, cisplatin and combine treatment group ( n = 5/group). J Comparison of tumor tissues from each group after experiment completion ( n = 5/group). K Immunohistochemical analysis results and scores of KI67 and PCNA ( n = 5/group). White bar means 100 μm. L HE staining results of major organs (heart, liver, gastrointestinal tract, lung, kidney) from each treatment group ( n = 5/group). Black bar means 100 μm.**, P < 0.01.

Article Snippet: The human immortalized hepatocyte cell line THLE-2, along with the liver cancer cell lines Huh1, Hep3B, JHH7, and Li-7, were sourced from Procell (Wuhan, China).

Techniques: Concentration Assay, Colony Assay, Double Staining, Flow Cytometry, In Vivo Imaging, Comparison, Immunohistochemical staining, Staining