Journal: Cell

Article Title: PEBP1 Wardens Ferroptosis by Enabling Lipoxygenase Generation of Lipid Death Signals

doi: 10.1016/j.cell.2017.09.044

Figure Lengend Snippet: KEY RESOURCE TABLE

Article Snippet: 15-LO2 Antibody (M-50), rabbit polyclonal IgG , Santa Cruz Biotechnology , Cat#sc-67143.

Techniques: Produced, Purification, Virus, Recombinant, Saline, Western Blot, Transfection, Protease Inhibitor, Membrane, Enzyme-linked Immunosorbent Assay, Binding Assay, Imaging, Software

Journal: Journal of Oncology

Article Title: SH3BGRL Suppresses Liver Tumor Progression through Enhanced ATG5-Dependent Autophagy

doi: 10.1155/2023/1105042

Figure Lengend Snippet: SH3BGRL arrests liver cell cycle progression: (a) flow cytometry analysis of cell cycle progression of liver cancer cells LO2 and HepG2 with SH3BGRL overexpression, compared with the control cells (Vect). Cells were grown at 50%–70% of confluency and starved for 12 h, then cultured with the full medium in less than 12 h. (b) Immunoblots of endogenous SH3BGRL by knockdown with two specific shSH3BGRL-1 and -2. (c) Flow cytometry of the indicated pool cells with SH3BGRL knockdown (shSH3BGRL-1,2) as well as their scramble infection control cells (Scr) as in (a). (d) Immunoblots of the indicated proteins in cells with SH3BGRL knockdown. Cells were cultured in either nutrient-sufficient or deficient conditions for 14 hours, followed by immunoblotting analysis.

Article Snippet: For knockdown of SH3BGRL in HepG2 and LO2 cells, two shRNA constructs against SH3BGRL (OriGene; Cat#TG309466) were transfected to establish HepG2 and LO2 SH3BGRL stable knockdown cell pools.

Techniques: Flow Cytometry, Over Expression, Control, Cell Culture, Western Blot, Knockdown, Infection

Journal: Journal of Oncology

Article Title: SH3BGRL Suppresses Liver Tumor Progression through Enhanced ATG5-Dependent Autophagy

doi: 10.1155/2023/1105042

Figure Lengend Snippet: SH3BGRL represses tumor formation of liver cells: (a) CCK-8 assay of the indicated cells with SH3BGRL knockdown under both nutrient-sufficient and deficient conditions. Data present as mean ± SD; n = 3; ∗∗ p < 0.01. (b, c) Xenograft tumor formation in nude mice by subcutaneous inoculation of 1 × 10 6 LO2 cells (b) or HepG2 cells (c) with SH3BGRL overexpression, along with their parental control cells (GFP). Cells were injected into the flanks of nude mice. After 28 days, mice were sacrificed and the formed tumor weight was scored. Mean ± SD; n = 5; ∗∗ p < 0.01; paired Student's t- test. (d, e) Xenograft tumor formation in nude mice by inoculation of either LO2 cells (d) or HepG2 cells (e) with SH3BGRL knockdown and the scrambled control (Scr) as mentioned above. Mean ± SD; n = 5; ∗∗ p < 0.01; unpaired Student's t- test.

Article Snippet: For knockdown of SH3BGRL in HepG2 and LO2 cells, two shRNA constructs against SH3BGRL (OriGene; Cat#TG309466) were transfected to establish HepG2 and LO2 SH3BGRL stable knockdown cell pools.

Techniques: CCK-8 Assay, Knockdown, Over Expression, Control, Injection

Journal: Journal of Oncology

Article Title: SH3BGRL Suppresses Liver Tumor Progression through Enhanced ATG5-Dependent Autophagy

doi: 10.1155/2023/1105042

Figure Lengend Snippet: SH3BGRL triggers basal autophagy of liver cells: (a, b) immunoblots of SH3BGRL and the autophagy-related proteins in cells with SH3BGRL overexpression (a) or endogenous SH3BGRL knockdown. β -actin was used as a loading control. GFP-vector (vector) and scrambled control (Scr) are shown. (c) Transmission electron microscopy of the formation of autophagosomes in LO2 and HepG2 cells with SH3BGRL overexpression. Enlarged images of the box are shown in the right panels. Representative images of double-membrane autophagosomes are pointed with arrows.

Article Snippet: For knockdown of SH3BGRL in HepG2 and LO2 cells, two shRNA constructs against SH3BGRL (OriGene; Cat#TG309466) were transfected to establish HepG2 and LO2 SH3BGRL stable knockdown cell pools.

Techniques: Western Blot, Over Expression, Knockdown, Control, Plasmid Preparation, Transmission Assay, Electron Microscopy, Membrane

Journal: Journal of Oncology

Article Title: SH3BGRL Suppresses Liver Tumor Progression through Enhanced ATG5-Dependent Autophagy

doi: 10.1155/2023/1105042

Figure Lengend Snippet: SH3BGRL colocalizes with the ATG5-ATG12 complex: (a, b) mutual immunoprecipitation of SH3BGRL with ATG5 in both LO2 and HepG2 cells with stable overexpression of SH3BGRL (a) or the parental normal cells with SH3BGRL and ATG5 antibodies, respectively. ATG5 and ATG12 were detected at their covalent complex position to indicate autophagy formation. (c) Immunofluorescence of the colocalization SH3BGRL with ATG5 in both LO2 and HepG2 cells with stable overexpression of SH3BGRL or the parental normal cells (GFP) with SH3BGRL and ATG5 antibodies, respectively. LC3 was stained to show its puncta to indicate the autophagy formation.

Article Snippet: For knockdown of SH3BGRL in HepG2 and LO2 cells, two shRNA constructs against SH3BGRL (OriGene; Cat#TG309466) were transfected to establish HepG2 and LO2 SH3BGRL stable knockdown cell pools.

Techniques: Immunoprecipitation, Over Expression, Immunofluorescence, Staining

Journal: Journal of Oncology

Article Title: SH3BGRL Suppresses Liver Tumor Progression through Enhanced ATG5-Dependent Autophagy

doi: 10.1155/2023/1105042

Figure Lengend Snippet: SH3BGRL stabilizes ATG5 from ubiquitination-mediated degradation: (a, b) immunoblots of ATG5 and ATG12 in LO2 cells with SH3BGRL overexpression or knockdown (a) or in HepG2 cells overexpressing SH3BGRL (b) with CHX or Act.D treatments. (c) Ubiquitination of ATG5 analysis by ATG5 antibody immunoprecipitation and immunoblotting in SH3BGRL-overexpressing HepG2 cells. MG132 was used to block ATG5 proteasome degradation.

Article Snippet: For knockdown of SH3BGRL in HepG2 and LO2 cells, two shRNA constructs against SH3BGRL (OriGene; Cat#TG309466) were transfected to establish HepG2 and LO2 SH3BGRL stable knockdown cell pools.

Techniques: Ubiquitin Proteomics, Western Blot, Over Expression, Knockdown, Immunoprecipitation, Blocking Assay

Journal: Journal of Oncology

Article Title: SH3BGRL Suppresses Liver Tumor Progression through Enhanced ATG5-Dependent Autophagy

doi: 10.1155/2023/1105042

Figure Lengend Snippet: ATG5 mediates SH3BGRL-induced tumor suppression: (a) CCK-8 assay of the indicated cells with additional ATG5 knockdown in SH3BGRL overexpression cells as indicated. Data present as mean ± SD; n = 3; ns: no significance; ∗∗ p < 0.01. (b) Immunoblots of the indicated proteins in SH3BGRL-overexpressing cells with additional ATG5 knockdown. The parental control cells (Vect) were used. (c) Xenograft tumor formation in nude mice by subcutaneous inoculation of HepG2 cells with additional ATG5 knockdown in SH3BGRL-overexpressing cells. Cells were injected into the flanks of nude mice. After 28 days, mice were sacrificed and the formed tumor weight was scored. Mean ± SD; n = 5; ∗∗ p < 0.01; paired Student's t- test.

Article Snippet: For knockdown of SH3BGRL in HepG2 and LO2 cells, two shRNA constructs against SH3BGRL (OriGene; Cat#TG309466) were transfected to establish HepG2 and LO2 SH3BGRL stable knockdown cell pools.

Techniques: CCK-8 Assay, Knockdown, Over Expression, Western Blot, Control, Injection

Journal: Journal of Oncology

Article Title: SH3BGRL Suppresses Liver Tumor Progression through Enhanced ATG5-Dependent Autophagy

doi: 10.1155/2023/1105042

Figure Lengend Snippet: Relevance of SH3BGRL-ATG5 autophagy in liver cancer: (a) immunoblots of SH3BGRL and the indicated autophagy-related proteins in fresh liver tumor samples (T) and their adjacent normal tissues (N). β -actin was used as a loading control. (b) Immunohistochemistry of SH3BGRL and ATG5 from The Protein Atlas database (details are indicated in Supplementary ). The bar is equal to 50 μ m. (c) Immunohistochemistry of SH3BGRL, LC3, and ATG5 in the xenograft mouse tumors in . Typical SH3BGRL-low and -high tumors are serially sliced. The bar is equal to 25 μ m. (d) Schematic model of SH3BGRL-ATG5-autophagy signaling in liver cancer repression.

Article Snippet: For knockdown of SH3BGRL in HepG2 and LO2 cells, two shRNA constructs against SH3BGRL (OriGene; Cat#TG309466) were transfected to establish HepG2 and LO2 SH3BGRL stable knockdown cell pools.

Techniques: Western Blot, Control, Immunohistochemistry

Journal: Journal of Gastrointestinal Oncology

Article Title: The deficiency of FKBP-5 inhibited hepatocellular progression by increasing the infiltration of distinct immune cells and inhibiting obesity-associated gut microbial metabolite

doi: 10.21037/jgo-21-71

Figure Lengend Snippet: FKBP-5 was highly expressed in HCC. (A) The expression of FKBP-5 was examined by the IHC staining of para-carcinoma tissues and HCC tissues. The representative images were displayed. Magnification, 100×; Scale bar =100 µm. (B) The western blotting analysis of FKBP-5 expression in HCC tissues (He) and para-carcinoma tissues (Pa), GAPDH was used as an internal reference. (C) The mRNA expression level of FKBP-5 was confirmed by RT-qPCR assay in HL-7702 and five HCC cells (SMMC-7721, Hep 3B, Huh7, Hep G2, and LO2), *, P<0.05; ***, P<0.001 vs. HL-7702 cells.

Article Snippet: Cell culture Human liver (HL)-7702 and HCC cells (i.e., SMMC-7721, Hep 3B, Huh7, Hep G2, and LO2) were purchased from the American Type Culture Collection.

Techniques: Expressing, Immunohistochemistry, Western Blot, Quantitative RT-PCR

Journal: Discovery Medicine

Article Title: Artesunate Inhibits Lipid Accumulation and Inflammation by Regulating the NLRP3 Inflammasome in Nonalcoholic Fatty Liver Disease

doi: 10.24976/discov.med.202436181.36

Figure Lengend Snippet: Fig. 1. Effect of artesunate (ART) on cell viability in LO2 cells after oleic acid (OA) treatment. (A) Effect of OA at different

Article Snippet: Human normal liver cells LO2 (TongPai, Shanghai, China) were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (CM10041, MACGENE, Beijing, China) supplemented with 10% fetal bovine serum (FBS, FCS500, EXcellBio, Shanghai, China) and 100 U/mL penicillin-streptomycin (P1400, Solarbio, Beijing, China).

Techniques:

Journal: Discovery Medicine

Article Title: Artesunate Inhibits Lipid Accumulation and Inflammation by Regulating the NLRP3 Inflammasome in Nonalcoholic Fatty Liver Disease

doi: 10.24976/discov.med.202436181.36

Figure Lengend Snippet: Fig. 2. ART restrained lipid accumulation in LO2 cells after OA treatment. (A) Lipid accumulation was demonstrated by oil red O staining (n = 3) (scale bar: 50 µm). (B) Effect of ART on triglycerides (TG) in LO2 cells after OA treatment (n = 3). (C) Effect of ART

Article Snippet: Human normal liver cells LO2 (TongPai, Shanghai, China) were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (CM10041, MACGENE, Beijing, China) supplemented with 10% fetal bovine serum (FBS, FCS500, EXcellBio, Shanghai, China) and 100 U/mL penicillin-streptomycin (P1400, Solarbio, Beijing, China).

Techniques: Staining

Journal: Discovery Medicine

Article Title: Artesunate Inhibits Lipid Accumulation and Inflammation by Regulating the NLRP3 Inflammasome in Nonalcoholic Fatty Liver Disease

doi: 10.24976/discov.med.202436181.36

Figure Lengend Snippet: Fig. 3. ART decreased the secretion of pro-inflammatory factors in OA treated LO2 cells. (A) Effect of ART on tumor necrosis

Article Snippet: Human normal liver cells LO2 (TongPai, Shanghai, China) were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (CM10041, MACGENE, Beijing, China) supplemented with 10% fetal bovine serum (FBS, FCS500, EXcellBio, Shanghai, China) and 100 U/mL penicillin-streptomycin (P1400, Solarbio, Beijing, China).

Techniques: