human normal hepatocytes lo2  (ATCC)

Journal: Translational Cancer Research

Article Title: Analysis of the role of POC1A in the development and progression of hepatocellular carcinoma

doi: 10.21037/tcr-23-2398

Figure Lengend Snippet: Analysis of the expression of POC1A in various classifications of HCC patients, as well as in HCC and liver cancer cell lines. (A) Comparison of POC1A between cancer tissue samples and normal tissue samples. (B) Comparison of the expression of POC1A in different genders. (C) Comparison of the expression of POC1A in different age groups. (D) The expression levels of POC1A were all higher in the Caucasian, African-American, and Asian HCC patients than the normal patients. The expression of POC1A was significantly higher in LIHC tissue samples from Asian patients than LIHC tissue samples from Caucasian patients. (E) The expression of POC1A in individual cancer stages. (F) The expression of POC1A in different transistor grades. (G) The expression of POC1A in tumor (T) stage. (H) The expression of POC1A in node (N) stage. (I) The expression of POC1A in metastasis (M) stage. (J) Analysis of POC1A expression in normal liver tissue and cancer tissue samples from patients with hepatocellular carcinoma using immunohistochemical methods in the Human Protein Atlas database. Liver sample: https://www.proteinatlas.org/ENSG00000164087-POC1A/tissue/liver#img , hepatocellular samples: https://www.proteinatlas.org/ENSG00000164087-POC1A/pathology/liver+cancer#img . Scale bar: 100 µm. (K) Analysis of POC1A mRNA expression in LO2 and HepG2 cell lines by qPCR assay. *, P<0.05; **, P<0.01; -: no significance. TCGA, The Cancer Genome Atlas; HCC, hepatocellular carcinoma; LIHC, liver hepatocellular carcinoma; mRNA, messenger RNA; qPCR, quantitative polymerase chain reaction; POC1A , POC1 centriolar protein A.

Article Snippet: The human normal hepatocyte cell line LO2 (Servicebio Technology Co., Ltd., Wuhan, China) was cultured in Roswell Park Memorial Institute 1640 medium, and the hepatoma cell line HepG2 (Procell Life Science & Technology Co., Ltd., Wuhan, China) was cultured in Dulbecco’s Modified Eagle Medium (Hyclone, Thermo Fisher Scientific Inc., Waltham, MA), supplemented with 10% fetal bovine serum and antibiotics (10,000 U/mL penicillin and 10 mg/mL streptomycin) (Procell Life Science & Technology Co., Ltd.).

Techniques: Expressing, Comparison, Immunohistochemical staining, Real-time Polymerase Chain Reaction

Journal: Frontiers in Pharmacology

Article Title: Ponatinib modulates the metabolic profile of obese mice by inhibiting adipose tissue macrophage inflammation

doi: 10.3389/fphar.2022.1040999

Figure Lengend Snippet: The metabolic profile of hepatocytes and adipocytes was not directly affected by ponatinib. (A) Representative Oil Red O staining of LO2 cells administrated with ponatinib or vehicle after FFA or BSA stimulation for 24 h n = 6. Scale bar, 20 μm (B) Quantitative analysis of oil red area of LO2 cells in (A) . Statistical comparisons were performed with one-way ANOVA followed by Bonferroni’s multiple comparisons post hoc test. Data represent mean ± SEM, *** p < 0.001 vs. BSA + vehical; n. s. p > 0.05 vs. FFA + Vehicle. (C) Representative Oil Red O staining of 3T3-L1 adipocytes treated with ponatinib or vehicle. n = 6. Scale bar, 200 μm (D) Quantitative analysis of oil red area of 3T3-L1 adipocytes in (C) . n = 6. Statistical comparisons were performed with unpaired two-tailed Student’s t-test, Data represent mean ± SEM, n. s. p > 0.05 vs . Vehicle. (E) Representative western blot showing levels of total and phosphorylation of IR-β (Tyr1150/1151), IRS1(Ser636/639), AKT (Ser473) of 3T3-L1 adipocytes in response to insulin stimulation for 30 min with or without ponatinib treatment. n = 4. (F–H) Quantification of phosphorylation of IR-β (Tyr1150/1151), IRS1 (Ser636/639), AKT (Ser473) expression level in (E) . n = 4. Statistical comparisons were performed with one-way ANOVA followed by Bonferroni’s multiple comparisons post hoc test. Data represent mean ± SEM, *** p < 0.001 vs. Vehical; n. s. p > 0.05 vs. Vehicle + Insulin. (I) The expression profile of ponatinib’s high-affinity targets in liver, skeletal muscle and omentum adipose from Genotype-Tissue Expression (GTEx) Project. Color scale indicates mean TPM (transcripts per million) value. (J) Representative electropherogram of one of six mice qPCR products to verify FGR, HCK, Lyn, CSFR, Fyn, FGFR, and ABL mRNA expression levels in BMDM, visceral fat, liver, and skeletal muscle.

Article Snippet: Human normal hepatocyte LO2 and mouse embryonic fibroblast 3T3-L1 were purchased from Procell Life Science and Technology Co. Ltd. All cell lines were maintained in standard medium containing Dulbecco’s modified Eagle’s medium (DMEM), 10% fetal bovine serum (FBS),1% penicillin–streptomycin at 37°C in 5% CO2.

Techniques: Staining, Two Tailed Test, Western Blot, Phospho-proteomics, Expressing