human hepatoma cell lines hepg2  (ATCC)

Journal: Tumour Virus Research

Article Title: PRMT5–mediated symmetric dimethylation of SHBs at Arg169 stabilizes SHBs and promotes angiogenesis and tumor growth

doi: 10.1016/j.tvr.2026.200340

Figure Lengend Snippet: SHBs is symmetrically dimethylated at arginine 169. (A–C) Huh7 and HepG2 cells were transfected with plasmids encoding SHBs–Strep–Flag or Strep–Flag control. Strep pull–down (IP:Strep) was performed, followed by Western blot (WB) with antibodies against (A) monomethylarginine (MMA), (B) asymmetric dimethylarginine (ADMA), or (C) symmetric dimethylarginine (SDMA). SHBs in the IP fraction and SHBs/β–actin in input lysates are shown as controls. (D) Cells expressing SHBs–Strep–Flag were treated with adenosine dialdehyde (ADOX, 40 μM) for 36 h, followed by Strep pull–down and WB for SDMA and SHBs. Densitometric ratios (SDMA/IP–SHBs and SHBs/β–actin) are shown above/below the blots. (E) Huh7 cells were transfected with plasmids encoding SHBs–Strep–Flag or the indicated R→K mutants (R73K, R78K, R79K, R169K). SDMA on immunoprecipitated SHBs was assessed by Strep pull–down and WB; densitometric SDMA/IP–SHBs ratios are shown above the blots. (F–G) HepG2 cells were transfected with plasmids encoding SHBs–Strep–Flag or SHBs/R169K–Strep–Flag (F) and SHBs/R169A–Strep–Flag (G) and analyzed by Strep pull–down and WB as in (E).

Article Snippet: Human hepatoma cell lines HepG2 (ATCC, HB–8065) and Huh7 (JCRB, JCRB0403), endothelial cell line EA.hy926 (ATCC, CRL–2922TM), and HEK293T cells (ATCC, CRL–3216) were obtained from the American Type Culture Collection (ATCC) and the Japanese Collection of Research Bioresources Cell Bank (JCRB, Japan).

Techniques: Transfection, Control, Western Blot, Expressing, Immunoprecipitation

Journal: Tumour Virus Research

Article Title: PRMT5–mediated symmetric dimethylation of SHBs at Arg169 stabilizes SHBs and promotes angiogenesis and tumor growth

doi: 10.1016/j.tvr.2026.200340

Figure Lengend Snippet: PRMT interacts with SHBs. (A) Huh7 cells were co–transfected with plasmids encoding SHBs–Strep–Flag (or Strep–Flag control) together with Flag–PRMT9. Strep pull–down was followed by WB with anti–Flag and anti–SHBs to assess co–precipitation. (B) Huh7 and HepG2 cells were co–transfected with plasmids encoding SHBs–Strep–Flag (or Strep–Flag control) together with Flag–PRMT5 and analyzed by Strep pull–down and WB as in (A). (C) Huh7 and HepG2 cells were co–transfected with plasmids encoding Strep–Flag–PRMT5 and SHBs–myc. Strep pull–down was performed and precipitates were immunoblotted for SHBs and Flag to validate the interaction. (D) Direct interaction between SHBs and PRMT5 was tested by GST pull–down. Purified GST or GST–PRMT5 (Coomassie–stained gel, left) was incubated with in vitro–translated SHBs–Flag, and bound SHBs was detected by WB using anti–Flag (right). (E) Confocal microscopy showing subcellular localization of SHBs (red) and PRMT5 (green) with nuclear DAPI staining (blue). Merged images and a representative line–scan fluorescence intensity profile (right) are shown.

Article Snippet: Human hepatoma cell lines HepG2 (ATCC, HB–8065) and Huh7 (JCRB, JCRB0403), endothelial cell line EA.hy926 (ATCC, CRL–2922TM), and HEK293T cells (ATCC, CRL–3216) were obtained from the American Type Culture Collection (ATCC) and the Japanese Collection of Research Bioresources Cell Bank (JCRB, Japan).

Techniques: Transfection, Control, Purification, Staining, Incubation, In Vitro, Confocal Microscopy, Fluorescence

Journal: Tumour Virus Research

Article Title: PRMT5–mediated symmetric dimethylation of SHBs at Arg169 stabilizes SHBs and promotes angiogenesis and tumor growth

doi: 10.1016/j.tvr.2026.200340

Figure Lengend Snippet: PRMT5 stabilizes SHBs protein expression in an Arg169–dependent manner. (A) Huh7 and HepG2 cells were co–transfected with plasmids encoding SHBs–Strep–Flag or SHBs/R169K–Strep–Flag together with increasing amounts of Flag–PRMT5 (0, 1, 3 μg). Whole–cell lysates were immunoblotted for SHBs, Flag, and β–actin; SHBs/β–actin ratios are shown above the blots. (B) Cells expressing SHBs–Strep–Flag or SHBs/R169K–Strep–Flag were transfected with NC or PRMT5 siRNAs (#1, #2). Lysates were immunoblotted for SHBs, PRMT5, and β–actin; SHBs/β–actin ratios are shown. (C–D) Cycloheximide (CHX) chase assays in (C) Huh7 and (D) HepG2 cells. Cells expressing SHBs or SHBs/R169K with vector or Flag–PRMT5 were treated with CHX for the indicated times (0–120 min), followed by WB for SHBs, Flag, and β–actin. Plots show relative SHBs levels normalized to time 0 with fitted linear regression (equations displayed). (E) HepG2 cells were co–transfected with plasmids encoding SHBs–Strep, HA–K48Ub, together with or without Flag–PRMT5, and treated with MG132 (20 μM) for 8 h, the ubiquitination levels of SHBs was evaluated via ubiquitination assay analysis. (F) HepG2 cells were co–transfected with plasmid encoding SHBs–Strep and TRIM21–myc (or control vector) and Flag–PRMT5 (or control vector), the cell lysates were subjected to immunoprecipitation using Strep–Tactin and analyzed by immunoblotting.

Article Snippet: Human hepatoma cell lines HepG2 (ATCC, HB–8065) and Huh7 (JCRB, JCRB0403), endothelial cell line EA.hy926 (ATCC, CRL–2922TM), and HEK293T cells (ATCC, CRL–3216) were obtained from the American Type Culture Collection (ATCC) and the Japanese Collection of Research Bioresources Cell Bank (JCRB, Japan).

Techniques: Expressing, Transfection, Plasmid Preparation, Ubiquitin Proteomics, Control, Immunoprecipitation, Western Blot

Journal: Tumour Virus Research

Article Title: PRMT5–mediated symmetric dimethylation of SHBs at Arg169 stabilizes SHBs and promotes angiogenesis and tumor growth

doi: 10.1016/j.tvr.2026.200340

Figure Lengend Snippet: Arg169 symmetric dimethylation is required for SHBs–driven angiogenesis and tumor growth. (A) WB analysis of SHBs and BIP expression in stably transduced Huh7 and HepG2 cells (Vector, SHBs, and SHBs/R169K). (B) ELISA measurement of VEGFA levels in the supernatants of Huh7/HepG2–Vector, Huh7/HepG2–SHBs, or Huh7/HepG2–SHBs/R169K cells. (C) Endothelial tube formation assay. EA.hy926 cells were cultured with conditioned media (CM) from Huh7 or HepG2 stable lines (Vector, SHBs, SHBs/R169K). Representative images and quantification of mesh numbers are shown. (D) Transwell migration assay. EA.hy926 cells were assessed for migration in response to CM from the indicated stable lines. Representative images and quantification of migrated cell numbers per field are shown. (E) Representative images of excised subcutaneous xenograft tumors derived from Huh7–Vector, Huh7–SHBs, or Huh7–SHBs/R169K cells. (F) Tumor growth curves (tumor volume over time) for the indicated xenograft groups. (G) Tumor weights at endpoint. (H) Representative immunohistochemical staining of xenograft tumors for CD31 and SHBs, with quantification of microvessel density (MVD) based on CD31 staining. Data are presented as mean ± SD; ∗ P < 0.05 as indicated.

Article Snippet: Human hepatoma cell lines HepG2 (ATCC, HB–8065) and Huh7 (JCRB, JCRB0403), endothelial cell line EA.hy926 (ATCC, CRL–2922TM), and HEK293T cells (ATCC, CRL–3216) were obtained from the American Type Culture Collection (ATCC) and the Japanese Collection of Research Bioresources Cell Bank (JCRB, Japan).

Techniques: Expressing, Stable Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Endothelial Tube Formation Assay, Cell Culture, Transwell Migration Assay, Migration, Derivative Assay, Immunohistochemical staining, Staining

Journal: Food Chemistry: Molecular Sciences

Article Title: Sangyod rice extract attenuates oleic acid–induced hepatic steatosis by modulating apoptotic, inflammatory, and lipid metabolic pathways

doi: 10.1016/j.fochms.2026.100387

Figure Lengend Snippet: Sangyod rice extract demonstrated a reduction in cytotoxicity and ROS levels in OA-induced HepG2 cells. (A) Viability of HepG2 cells exposed to different concentrations of Sangyod rice extract. (B) Viability of Sangyod rice extract treatment after OA-induced HepG2 cells. (C) ROS generation in OA-induced HepG2 cells. Results are presented as the mean ± SEM from four independent biological experiments ( n = 4). One-way ANOVA followed by Tukey ' s post hoc test was used to determine statistical significance. * p < 0.05 compared to the control group, and # p < 0.05 compared to the OA group. Groups: Control (0.1% DMSO); OA (0.4 mM), oleic acid-induced HepG2 cells without treatment; SR 10, OA-induced HepG2 cells +10 μg/mL Sangyod rice extract; SR 50, OA-induced HepG2 cells +50 μg/mL Sangyod rice extract; SR 100, OA-induced HepG2 cells +100 μg/mL Sangyod rice extract.

Article Snippet: The HepG2 human hepatocellular carcinoma cell line was procured from the American Type Culture Collection (Manassas, VA, USA) and nurtured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) enriched with 10% fetal bovine serum (Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (Gibco, Waltham, MA, USA), and 1% l -glutamine (Gibco, Waltham, MA, USA).

Techniques: Control

Journal: Food Chemistry: Molecular Sciences

Article Title: Sangyod rice extract attenuates oleic acid–induced hepatic steatosis by modulating apoptotic, inflammatory, and lipid metabolic pathways

doi: 10.1016/j.fochms.2026.100387

Figure Lengend Snippet: Sangyod rice extract inhibited apoptosis in OA-induced HepG2 cells by suppressing the Bax and caspase-3 pathway. (A) Representative images of nuclei stained with Hoechst 33342. Images shown at ×20 magnification. Scale bar: 50 μm. (B) Percentage of apoptotic cells after treatment with Sangyod rice extract in OA-induced HepG2 cells. (C) Western blot analysis of Bax, Bcl-2, procaspase-3, and cleaved caspase-3. (D) Relative expression of Bax and Bcl-2. (E) Relative expression of procaspase 3, and cleaved caspase 3. Results are presented as the mean ± SEM from four independent biological experiments ( n = 4). One-way ANOVA followed by Tukey ' s post hoc test was used to determine statistical significance. *p < 0.05 compared to the control group, and #p < 0.05 compared to the OA group. Groups: Control (0.1% DMSO); OA (0.4 mM), oleic acid-induced HepG2 cells without treatment; SR 10, OA-induced HepG2 cells +10 μg/mL Sangyod rice extract; SR 50, OA-induced HepG2 cells +50 μg/mL Sangyod rice extract; SR 100, OA-induced HepG2 cells +100 μg/mL Sangyod rice extract.

Article Snippet: The HepG2 human hepatocellular carcinoma cell line was procured from the American Type Culture Collection (Manassas, VA, USA) and nurtured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) enriched with 10% fetal bovine serum (Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (Gibco, Waltham, MA, USA), and 1% l -glutamine (Gibco, Waltham, MA, USA).

Techniques: Staining, Western Blot, Expressing, Control

Journal: Food Chemistry: Molecular Sciences

Article Title: Sangyod rice extract attenuates oleic acid–induced hepatic steatosis by modulating apoptotic, inflammatory, and lipid metabolic pathways

doi: 10.1016/j.fochms.2026.100387

Figure Lengend Snippet: Sangyod rice extract attenuated inflammation in OA-induced HepG2 cells through inhibition of the NF-κB pathway. (A) TNF-α gene, (B) IL-1β gene, (C) IL-6 gene, (D) IL-10 gene. (E) Western blot analysis of NF-κB. (F) Relative expression of NF-κB protein. Results are presented as the mean ± SEM from four independent biological experiments (n = 4). One-way ANOVA followed by Tukey ' s post hoc test was used to determine statistical significance. *p < 0.05 indicates significance compared to the control group, while #p < 0.05 denotes significance compared to the OA group. Groups: Control (0.1% DMSO); OA (0.4 mM), oleic acid-induced HepG2 cells without treatment; SR 10, OA-induced HepG2 cells +10 μg/mL Sangyod rice extract; SR 50, OA-induced HepG2 cells +50 μg/mL Sangyod rice extract; SR 100, OA-induced HepG2 cells +100 μg/mL Sangyod rice extract.

Article Snippet: The HepG2 human hepatocellular carcinoma cell line was procured from the American Type Culture Collection (Manassas, VA, USA) and nurtured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) enriched with 10% fetal bovine serum (Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (Gibco, Waltham, MA, USA), and 1% l -glutamine (Gibco, Waltham, MA, USA).

Techniques: Inhibition, Western Blot, Expressing, Control

Journal: Food Chemistry: Molecular Sciences

Article Title: Sangyod rice extract attenuates oleic acid–induced hepatic steatosis by modulating apoptotic, inflammatory, and lipid metabolic pathways

doi: 10.1016/j.fochms.2026.100387

Figure Lengend Snippet: Sangyod rice extract reduced lipid accumulation in OA-induced HepG2 cells. (A) Oil Red O staining was conducted on HepG2 cells, with red fat droplets indicating lipid accumulation. Images shown at ×20 magnification. Scale bar: 50 μm. (B) Percentage of lipid accumulation post Oil Red O extraction. (C) Levels of TG were measured using an assay kit. The data is displayed as mean ± SEM from four independent biological experiments (n = 4). One-way ANOVA followed by Tukey ' s post hoc test was used to determine statistical significance. *p < 0.05 indicates significance compared to the control group, while #p < 0.05 denotes significance compared to the OA group. Groups: Control (0.1% DMSO); OA (0.4 mM), oleic acid-induced HepG2 cells without treatment; SR 10, OA-induced HepG2 cells +10 μg/mL Sangyod rice extract; SR 50, OA-induced HepG2 cells +50 μg/mL Sangyod rice extract; SR 100, OA-induced HepG2 cells +100 μg/mL Sangyod rice extract.

Article Snippet: The HepG2 human hepatocellular carcinoma cell line was procured from the American Type Culture Collection (Manassas, VA, USA) and nurtured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) enriched with 10% fetal bovine serum (Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (Gibco, Waltham, MA, USA), and 1% l -glutamine (Gibco, Waltham, MA, USA).

Techniques: Staining, Extraction, Control

Journal: Food Chemistry: Molecular Sciences

Article Title: Sangyod rice extract attenuates oleic acid–induced hepatic steatosis by modulating apoptotic, inflammatory, and lipid metabolic pathways

doi: 10.1016/j.fochms.2026.100387

Figure Lengend Snippet: Effect of Sangyod rice extract on lipid metabolism in OA-induced HepG2 cells. (A) SREBP-1c gene (B) ACC gene, (C) FASN gene (D) CPT-1 A gene, (E) SCD1 gene, (F) MTTP gene. The data is displayed as mean ± SEM from four independent biological experiments (n = 4). One-way ANOVA followed by Tukey ' s post hoc test was used to determine statistical significance. *p < 0.05 indicates significance compared to the control group, while #p < 0.05 denotes significance compared to the OA group. Groups: Control (0.1% DMSO); OA (0.4 mM), oleic acid-induced HepG2 cells without treatment; SR 10, OA-induced HepG2 cells +10 μg/mL Sangyod rice extract; SR 50, OA-induced HepG2 cells +50 μg/mL Sangyod rice extract; SR 100, OA-induced HepG2 cells +100 μg/mL Sangyod rice extract.

Article Snippet: The HepG2 human hepatocellular carcinoma cell line was procured from the American Type Culture Collection (Manassas, VA, USA) and nurtured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) enriched with 10% fetal bovine serum (Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (Gibco, Waltham, MA, USA), and 1% l -glutamine (Gibco, Waltham, MA, USA).

Techniques: Control

Journal: Food Chemistry: Molecular Sciences

Article Title: Sangyod rice extract attenuates oleic acid–induced hepatic steatosis by modulating apoptotic, inflammatory, and lipid metabolic pathways

doi: 10.1016/j.fochms.2026.100387

Figure Lengend Snippet: Effect of Sangyod rice extract on the expression of LPL-1, LPL-2, PGC-1α and PPARα in OA-induced HepG2 cells. (A) LPL-1 gene (B) LPL-2 gene, (C) PPARα gene (D) PGC-1α gene. The data is displayed as mean ± SEM from four independent biological experiments (n = 4). One-way ANOVA followed by Tukey ' s post hoc test was used to determine statistical significance. *p < 0.05 indicates significance compared to the control group, while #p < 0.05 denotes significance compared to the OA group. Groups: Control (0.1% DMSO); OA (0.4 mM), oleic acid-induced HepG2 cells without treatment; SR 10, OA-induced HepG2 cells +10 μg/mL Sangyod rice extract; SR 50, OA-induced HepG2 cells +50 μg/mL Sangyod rice extract; SR 100, OA-induced HepG2 cells +100 μg/mL Sangyod rice extract.

Article Snippet: The HepG2 human hepatocellular carcinoma cell line was procured from the American Type Culture Collection (Manassas, VA, USA) and nurtured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) enriched with 10% fetal bovine serum (Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (Gibco, Waltham, MA, USA), and 1% l -glutamine (Gibco, Waltham, MA, USA).

Techniques: Expressing, Control

Journal: Food Chemistry: Molecular Sciences

Article Title: Sangyod rice extract attenuates oleic acid–induced hepatic steatosis by modulating apoptotic, inflammatory, and lipid metabolic pathways

doi: 10.1016/j.fochms.2026.100387

Figure Lengend Snippet: Sangyod rice extract regulates lipid metabolism through the Akt and MAPK signaling pathways. (A) Western blot analysis of Akt, ERK1/2 amd p38 MAPK, (B) Relative expression of pERK/ERK protein, (C) Relative expression of p-p38/p38 protein, (D) Relative expression of pAkt/Akt protein. The data is displayed as mean ± SEM from four independent biological experiments (n = 4). One-way ANOVA followed by Tukey ' s post hoc test was used to determine statistical significance. *p < 0.05 indicates significance compared to the control group, while #p < 0.05 denotes significance compared to the OA group. Groups: Control (0.1% DMSO); OA (0.4 mM), oleic acid-induced HepG2 cells without treatment; SR 10, OA-induced HepG2 cells +10 μg/mL Sangyod rice extract; SR 50, OA-induced HepG2 cells +50 μg/mL Sangyod rice extract; SR 100, OA-induced HepG2 cells +100 μg/mL Sangyod rice extract.

Article Snippet: The HepG2 human hepatocellular carcinoma cell line was procured from the American Type Culture Collection (Manassas, VA, USA) and nurtured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) enriched with 10% fetal bovine serum (Gibco, Waltham, MA, USA), 1% penicillin/streptomycin (Gibco, Waltham, MA, USA), and 1% l -glutamine (Gibco, Waltham, MA, USA).

Techniques: Protein-Protein interactions, Western Blot, Expressing, Control

Journal: bioRxiv

Article Title: Phosphoglycerate mutase 5 regulates lipid metabolism and mitochondrial homeostasis in hepatocellular cancer cells

doi: 10.64898/2026.05.01.718031

Figure Lengend Snippet: a . Representative images of PGAM5 (green) immunohistochemical labeling in canine normal liver, peritumoral liver, and HCC. Nuclei are stained with Hoechst (blue). b . Quantification of immunohistochemical PGAM5 expression in dogs; data include normal liver (n = 12), peritumoral liver (n = 21), and HCC (n = 37). Data expressed as mean ± SEM of percentage of PGAM5-positive cells. c. Images of mitochondria (MitoTracker, GreenFM), superoxide (MitoSOX, Red), and acidic lysosomes (LysoTracker, Blue) in WT and PGAM5 KO Huh7 cells 30-60 minutes after exposure to nutrient depleted media. d. e. Fluorometric assay measurement of ROS in WT and PGAM5 KO cells after 16-hour exposure to complete or depleted culture media in Huh7 (d) and HepG2 (e) cells. Data expressed as mean ± SEM. f. Violin plots of mitochondrial DNA (mtDNA) copy number in WT and PGAM5 KO Huh7 and HepG2 cells. g., h. Real-time (RT) qPCR of Huh7 (g.) and HepG2 (h.) wild type (black square) and PGAM5 KO (grey triangle) PGC1a transcripts. Data represent 3 biological replicates with a minimum of 2 technical replicates per sample. Data expressed as mean ± SEM; *, p<0.05.

Article Snippet: The HepG2 (HB-8065) hepatoma cell line was purchased from ATCC and maintained in EMEM media (30-2003, ATCC) supplemented with 10% FBS and 1% PS.

Techniques: Immunohistochemical staining, Labeling, Staining, Expressing, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Phosphoglycerate mutase 5 regulates lipid metabolism and mitochondrial homeostasis in hepatocellular cancer cells

doi: 10.64898/2026.05.01.718031

Figure Lengend Snippet: a . Graph of oxygen consumption rate (OCR) in wild type (WT, black squares) and PGAM5 knockout (PGAM5 KO ; grey triangles) Huh7 cells. Displayed data are derived from 3 biological replicates with a minimum of 3 technical replicates per sample. Results are normalized to cell count. b . Graph of extracellular acidification rate (ECAR) in WT (black squares) and PGAM5 KO (grey triangles) Huh7 cells. c. Schematic of the Agilent Seahorse fatty acid fuel flex assay. d. Dependency percentage and capacity percentage of fatty acids, glucose, and glutamine in WT (black circles) and PGAM5 KO (grey triangles) Huh7 cells. Displayed fuel flex data are derived from 3 biological replicates with a minimum of 3 technical replicates per sample. e. Acetyl-CoA concentration in WT and PGAM5KO HepG2 and Huh7 cells. Data represent 3 biological replicates with a minimum of 2 technical replicates per sample f. Representative transmission electron micrographs of Huh7 WT and PGAM5 KO treated with 500 uM palmitate for 16 hours. Black ovals encircle mitochondria, “L” labels lipid droplets, and scale bars equal 2.0 µm. g. Isotope-Ratio Mass Spectrometry atom percentage (%) excess of 13 C in Huh7 WT and PGAM5 KO pelleted cells or supernatant following treatment with 500uM 13 C-palmitate (PA) or unlabeled bovine serum albumin (BSA) for 16 hours. Data represent 3 biological replicates with 2 technical replicates per sample; *, p<0.05.

Article Snippet: The HepG2 (HB-8065) hepatoma cell line was purchased from ATCC and maintained in EMEM media (30-2003, ATCC) supplemented with 10% FBS and 1% PS.

Techniques: Knock-Out, Derivative Assay, Cell Characterization, Concentration Assay, Transmission Assay, Mass Spectrometry

Journal: bioRxiv

Article Title: Phosphoglycerate mutase 5 regulates lipid metabolism and mitochondrial homeostasis in hepatocellular cancer cells

doi: 10.64898/2026.05.01.718031

Figure Lengend Snippet: a. Top 20 downregulated genes (red) and upregulated (Blue) genes secondary to PGAM5 deletion in Huh7 cells. b . Gene ontologies (GO) plot of enriched biological processes (BP, orange), cellular components (CC, green), and molecular functions (MF, blue) associated with PGAM5 expression in Huh7 cells. c., d. STRING analyses of Huh7 ( c ) and ( d ) HepG2 GO regulation of lipid metabolism (GO:0019216). PPI enrichment p-value 1.06e-10, FDR < 0.02. e., f. Real-time (RT) qPCR of HepG2 ( e ) and Huh7 ( f ) wild type (black square) and PGAM5 KO (grey triangle) of fatty acid synthase ( FASN ). Data are derived from a minimum of 2 biological replicates with 2 technical replicates per sample and are represented as mean ± SEM; *, p<0.05. g. Representative immunoblots of acetyl-CoA carboxylase (ACC), FASN, and actin from WT and PGAM5 KO HepG2 and Huh7 cell lysates. Relative protein expression is normalized to actin. Data represent 3 biological replicates presented as mean ± SD; *, p<0.05.

Article Snippet: The HepG2 (HB-8065) hepatoma cell line was purchased from ATCC and maintained in EMEM media (30-2003, ATCC) supplemented with 10% FBS and 1% PS.

Techniques: Expressing, Quantitative RT-PCR, Derivative Assay, Western Blot

Journal: bioRxiv

Article Title: Phosphoglycerate mutase 5 regulates lipid metabolism and mitochondrial homeostasis in hepatocellular cancer cells

doi: 10.64898/2026.05.01.718031

Figure Lengend Snippet: a. Comparative derived total abundance of diacylglycerol (DAG) in the Huh7 lipidome of wildtype (WT) and PGAM5 knockout (KO) cells, mean ± SD. b. c. Relative quantification of diacylglycerol (DAG) abundance in wild type (WT) and PGAM5 KO HepG2 (f) and Huh7 (g) cells cultured with bovine serum albumin (BSA, filled circles) or palmitic acid (PA, open triangles). AU - arbitrary units. Displayed data are derived from 3 biological replicates with a minimum of 2 technical replicates per sample. d. Coomassie blue stained PhosTag gel of lipin-1 protein immunoprecipitated from Huh7 WT and PGAM5 KO Huh7 cells. e. Proposed mechanistic link: PGAM5 mediated dephosphorylation of lipin-1 modulates metabolic activity by upregulating DAG synthesis from phosphatidic acid and promoting transcription of genes regulating beta-oxidation. f.-i. Real-time (RT) qPCR relative expression of Huh7 ( f, h ) and HepG2 ( g.i ) wild type (black square) and PGAM5 KO (grey triangle) transcripts fatty acid binding protein 1 ( FABP1 , f, g ) and carnitine palmitoyltransferase 1A ( CPT1a , h, i ). RT qPCR data are derived from a minimum of 3 biological replicates with 2 technical replicates per sample, mean ± SEM; *, p<.05.

Article Snippet: The HepG2 (HB-8065) hepatoma cell line was purchased from ATCC and maintained in EMEM media (30-2003, ATCC) supplemented with 10% FBS and 1% PS.

Techniques: Derivative Assay, Knock-Out, Quantitative Proteomics, Cell Culture, Staining, Immunoprecipitation, De-Phosphorylation Assay, Activity Assay, Quantitative RT-PCR, Expressing, Binding Assay

Journal: Nature Communications

Article Title: Charting spatial ligand-target activity using Renoir

doi: 10.1038/s41467-026-72388-7

Figure Lengend Snippet: a Ranking of activity of ligands expressed by onco-fetal cell types with other major cell types present in the neighborhoods of the onco-fetal cells for the CosMx hepatocellular carcinoma (HCC) dataset. Stacked color bars represent the cell types that express the ligand (left) and target (bottom). b Distribution of ligand-target interactions across target cell types. The target cell types include major non-onco fetal cell types present in the neighborhoods of the onco-fetal cell types c (left) Spatial map of cell type specific neighborhood activity scores across cell types associated with the ligand and target for VEGFA:KDR and CXCL12:CXCR4 respectively (fov 10); (right) Spatial distribution of cell types associated with the ligand and target for VEGFA:KDR and CXCL12:CXCR4 respectively (fov 10). White scale bar, 125 μm. d (top) Spatial map of cell type specific neighborhood activity scores across cell types associated with the ligand and target for IL6:PTGS2 (fov 10); (bottom) Spatial distribution of cell types associated with the ligand and target for IL6:PTGS2 (fov 10) e IL6 induced bipotent stem cell-like properties in HepG2 cells. Treatment with IL6 induced CD133 + population in a dose-dependent manner. The percentage of CD133 + HepG2 cells was measured by flow cytometry after the HepG2 cells were treated with IL6 protein for increasing concentration for 96 h. All data presented are from three independent experiments containing three technical replicates ( n = 9). p -values (* p < 0.005 vs control groups; ** p < 0.0005 vs control groups) are calculated using two-sided Wilcoxon rank-sum test. p -values: 0.2ng = 0.051938, 2ng = 0.001987, 20 ng = 0.002666, 200 ng = 0.000409, 2 μg = 0.000409. The box denotes the interquartile range (IQR, the range between the 25 th and 75 th percentile) with the median value, whiskers indicate the maximum and minimum value within 1.5 times the IQR. Individual datapoints are overlaid on the boxplots. Created in BioRender. Hou, S. (2026) ( https://BioRender.com/7pacpt3 ). Source data are provided as a Source Data file.

Article Snippet: HepG2 cells line (ATCC, HB-8065) was cultured in DMEM (gibco, REF: 11995065) with 10% FBS (Garvan Institute of Medical Research) and 1% Penicillin Streptomycin (gibco, REF: 15070063).

Techniques: Activity Assay, Flow Cytometry, Concentration Assay, Control