normal hepatocyte cell line l02  (Genechem)

Journal: The Journal of Biological Chemistry

Article Title: G protein–coupled estrogen receptor 1 ameliorates nonalcoholic steatohepatitis through targeting AMPK-dependent signaling

doi: 10.1016/j.jbc.2024.105661

Figure Lengend Snippet: GPER1 plays a crucial role in the protective effects of estradiol on lipid accumulation, oxidative stress, and inflammation in L02 cells under metabolic stress. A , representative images of Nile Red staining in L02 cells treated with vehicle or GPER1-specific agonist G1 (100 nM) followed by BSA or PO (palmitic acid and oil acid mixture) stimulation for 12 h (n = 3 independent experiments). Scale bar represents 100 μm. B , triglyceride (TG) and total cholesterol (TC) contents in L02 cells from the indicated group (n = 6). C , relative mRNA levels of factors related to fatty acid metabolism in L02 cells from the indicated group (n = 6). D , representative images of DCFH-DA probe–stained L02 cells in the indicated group (n = 3 independent experiments). Scale bar represents 100 μm. E , representative images of MitoSOX Red probe–stained L02 cells in the indicated group (n = 3 independent experiments). Scale bar represents 100 μm. F , relative mRNA levels of factors related to inflammatory response in L02 cells from the indicated group (n = 6). G , representative images of Nile Red staining of L02 cells challenged by PO and treated with vehicle, 17β-estradiol (E2; 10 nM), GPER1 antagonist G15 (10 μM), or E2 in combination with G15 (n = 3 independent experiments). Scale bar represents 100 μm. H , TG and TC contents in L02 cells from the indicated group (n = 6). I , relative mRNA levels of factors related to fatty acid metabolism in L02 cells from the indicated group (n = 6). J , representative images of DCFH-DA probe–stained L02 cells in the indicated group (n = 3 independent experiments). Scale bar represents 100 μm. K , representative images of MitoSOX Red probe–stained L02 cells in the indicated group (n = 3 independent experiments). Scale bar represents 100 μm. L , relative mRNA levels of factors related to inflammatory response in L02 cells from the indicated group (n = 6). M , immunoblotting analysis of GPER1 protein level in the WT and GPER1 KO L02 cells (n = 3 independent experiments). N , representative images of Nile Red staining in the WT and GPER1 KO L02 cells challenged by PO and cotreated with vehicle or E2 (n = 3 independent experiments). Scale bar represents 100 μm. O , TG and TC contents in L02 cells from the indicated group (n = 6). P , relative mRNA levels of factors related to fatty acid metabolism in L02 cells from the indicated group (n = 6). Q , representative images of DCFH-DA probe–stained L02 cells in the indicated group (n = 3 independent experiments). Scale bar represents 100 μm. R , representative images of MitoSOX Red probe–stained L02 cells in the indicated group (n = 3 independent experiments). Scale bar represents 100 μm. S , relative mRNA levels of factors related to inflammatory response in L02 cells from the indicated group (n = 6). In all statistical plots, data are expressed as the mean ± SD and analyzed by one-way ANOVA with Bonferroni analysis. ∗∗∗ p < 0.001, comparison between the indicated groups; n.s., no significance, p ≥ 0.05, comparison between the indicated groups. The mRNA expression of target genes was normalized to that of Actb. BSA, bovine serum albumin; GPER1, G protein–coupled estrogen receptor 1; PO, palmitic acid-oleic acid mixture.

Article Snippet: The L02 cell lines (human normal hepatocyte) were purchased from the China Center for Type Culture Collection and cultured in Roswell Park Memorial Institute 1640 medium with 10% FBS and 1% penicillin-streptomycin.

Techniques: Staining, Western Blot, Comparison, Expressing

Journal: The Journal of Biological Chemistry

Article Title: G protein–coupled estrogen receptor 1 ameliorates nonalcoholic steatohepatitis through targeting AMPK-dependent signaling

doi: 10.1016/j.jbc.2024.105661

Figure Lengend Snippet: GPER1 mediates the release of cAMP and activates the AMPK signaling pathway. A , cyclic AMP levels in PO (palmitic acid and oil acid mixture)-challenged L02 cells for 30 min after treatment with 10 −10 to 10 −5 M GPER1-specific agonist G1 (n = 4). B , cyclic AMP levels in PO-challenged L02 cells after treatment with G1 (100 nM) for the indicated times (n = 4). C , cyclic AMP levels in WT or GPER1 KO L02 cells challenged by PO and cotreated with vehicle or 17β-estradiol (E2; 10 nM) for the indicated times (n = 3). D , cyclic AMP levels in WT or GPER1 KO HepG2 cells were challenged by PO and cotreated with vehicle or E2 for the indicated times (n = 3). E , immunoblotting analyses of total and phosphorylated AMPKα and ACCα protein levels in primary hepatocytes that isolate from WT female mice challenged by PO and cotreated with vehicle or G1 in the absence or presence of Gαs inhibitor NF449 (10 μM). F , immunoblotting analyses of total and phosphorylated AMPKα and ACCα protein levels in L02 cells ( left ) and primary hepatocytes ( right ) that isolate from WT female mice challenged by PO and cotreated with vehicle or G1 in the absence or presence of AC inhibitor MDL-12330A (20 μM). G , immunoblotting analyses of total and phosphorylated AMPKα and ACCα protein levels in L02 cells ( left ), HepG2 cells ( middle ), and primary hepatocytes ( right ) that isolate from WT female mice challenged by PO and cotreated with vehicle or G1 in the absence or presence of EPAC antagonist ESI-09 (10 μM). H , immunoblotting analyses of total and phosphorylated AMPKα and ACCα protein levels in L02 cells ( left ), HepG2 cells ( middle ), and primary hepatocytes ( right ) that isolate from WT female mice challenged by PO and cotreated with vehicle or G1 in the absence or presence of PKA inhibitor H89 (10 μM). I , immunoblotting analyses of total and phosphorylated AMPKα and ACCα protein levels in L02 cells ( left ) and HepG2 cells ( right ) challenged by PO and cotreated with vehicle or G1 in the absence or presence of PKA catalytic subunit α (PKAc) siRNA. In all statistical plots, data are expressed as the mean ± SD and analyzed by one-way ANOVA with Bonferroni analysis. For ( A ) and ( B ), ∗ p < 0.05, ∗∗∗ p < 0.001, G1-PO group versus Vehicle-PO group. For ( C ) and ( D ), ∗∗∗ p < 0.001, comparison between the indicated groups; n.s., no significance, p ≥ 0.05, comparison between the indicated groups. AC, adenylyl cyclase; AMPK, AMP-activated protein kinase; EPAC, exchange protein activated by cAMP; GPER1, G protein–coupled estrogen receptor 1; PO, palmitic acid-oleic acid mixture.

Article Snippet: The L02 cell lines (human normal hepatocyte) were purchased from the China Center for Type Culture Collection and cultured in Roswell Park Memorial Institute 1640 medium with 10% FBS and 1% penicillin-streptomycin.

Techniques: Western Blot, Comparison

Journal: American Journal of Cancer Research

Article Title: The effect of hepatocellular carcinoma-associated fibroblasts on hepatoma vasculogenic mimicry

doi:

Figure Lengend Snippet: Observing the ability of vasculogenic mimicry in different cell lines in three-dimensional culture. A. L02; B. SMMC-7721; C. MHCC-97H. Bar=100 μm.

Article Snippet: The human normal hepatocyte cell line L02, human hepatoma cell line SMMC-7721 and MHCC-97H were purchased from Beijing Zhongshan Jinqiao Biotechnology Co., Ltd. L02 cells were cultured in RPMI-1640 medium containing 10% FBS.

Techniques: