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: iScience

Article Title: F-53B exposure accelerates progression from preexisting fatty liver to non-alcoholic steatohepatitis and hepatic fibrosis

doi: 10.1016/j.isci.2026.115675

Figure Lengend Snippet: Knockdown of L-FABP mitigated F-53B-induced damage in HepG2 cells (A) Liver-type fatty acid-binding protein (L-FABP) knockdown was confirmed by immunoblot analysis using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as the loading control ( n = 6). (B) Calculated L-FABP/GAPDH ratio ( n = 6) based on the immunoblot results shown in (A). (C) Effects of different treatments on TG content in HepG2 cells ( n = 5). (D) Fold change of TG content in NC + F-53B/NC and L-FABP KD + F-53B/L-FABP KD groups ( n = 5). (E) Interleukin-6 (IL-6) expression in HepG2 cells following different treatments. (F) Relative IL-6 expression (IL-6/GAPDH) ( n = 6). (G) Fold change of IL-6 (relative to respective control) in NC vs. L-FABP KD groups with F-53B treatment ( n = 6). (H) Transforming growth factor β1 (TGF-β1) expression in HepG2 cells following different treatments. (I) Relative TGF-β1 expression (TGF-β1/GAPDH) ( n = 6). (J) Fold change of TGF-β1 (relative to respective control) in NC vs. L-FABP KD groups with F-53B treatment ( n = 6). NC, negative control siRNA group. NC + F-53B, NC with 5 mg/L F-53B group. L-FABP KD, L-FABP-knockdown group. L-FABP KD + F-53B, L-FABP KD with 5 mg/L F-53B group. Data are presented as mean ± SEM. The significance of differences between two groups was determined using the Mann-Whitney U test, and is indicated by hash symbols: # p < 0.05, ## p < 0.01.

Article Snippet: HepG2 cells were authenticated by ATCC using STR profiling and tested negative for mycoplasma contamination.

Techniques: Knockdown, Binding Assay, Western Blot, Control, Expressing, Negative Control, MANN-WHITNEY

Journal: iScience

Article Title: F-53B exposure accelerates progression from preexisting fatty liver to non-alcoholic steatohepatitis and hepatic fibrosis

doi: 10.1016/j.isci.2026.115675

Figure Lengend Snippet: Knockdown of L-FABP mitigated F-53B-induced damage in HepG2 cells (A) Liver-type fatty acid-binding protein (L-FABP) knockdown was confirmed by immunoblot analysis using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as the loading control ( n = 6). (B) Calculated L-FABP/GAPDH ratio ( n = 6) based on the immunoblot results shown in (A). (C) Effects of different treatments on TG content in HepG2 cells ( n = 5). (D) Fold change of TG content in NC + F-53B/NC and L-FABP KD + F-53B/L-FABP KD groups ( n = 5). (E) Interleukin-6 (IL-6) expression in HepG2 cells following different treatments. (F) Relative IL-6 expression (IL-6/GAPDH) ( n = 6). (G) Fold change of IL-6 (relative to respective control) in NC vs. L-FABP KD groups with F-53B treatment ( n = 6). (H) Transforming growth factor β1 (TGF-β1) expression in HepG2 cells following different treatments. (I) Relative TGF-β1 expression (TGF-β1/GAPDH) ( n = 6). (J) Fold change of TGF-β1 (relative to respective control) in NC vs. L-FABP KD groups with F-53B treatment ( n = 6). NC, negative control siRNA group. NC + F-53B, NC with 5 mg/L F-53B group. L-FABP KD, L-FABP-knockdown group. L-FABP KD + F-53B, L-FABP KD with 5 mg/L F-53B group. Data are presented as mean ± SEM. The significance of differences between two groups was determined using the Mann-Whitney U test, and is indicated by hash symbols: # p < 0.05, ## p < 0.01.

Article Snippet: HepG2 human hepatocellular carcinoma cells (ATCC, HB-8065) were maintained in a humidified incubator at 37 °C with 5% CO 2 .

Techniques: Knockdown, Binding Assay, Western Blot, Control, Expressing, Negative Control, MANN-WHITNEY

Journal: PLOS One

Article Title: Unraveling the role of ChREBP in lung adenocarcinoma: Expression, regulatory networks, and potential functional impact

doi: 10.1371/journal.pone.0347907

Figure Lengend Snippet: (A) Expression levels of ChREBP ( MLXIPL ) in 66 human LUAD cell lines, derived from RNA-seq datasets available in the EMBL-EBI Expression Atlas, are presented in transcripts per million (TPM). (B-F) Expression of ChREBP in NCI-H1975, NCI-H1650, NCI-H2228 and HepG2 cells assessed using RT-qPCR assays. (B) Expression level of seven candidate HKGs. Cycle threshold (Ct) values are displayed. (C) Average expression stability values of seven HKGs determined by geNorm are shown. Expression levels of total ChREBP (D), ChREBP-α (E), and ChREBP-β (F) were normalized against the three most stable HKGs, RPS13, QARS, and RNA18S . Data represents the results of three independent experiments. Asterisks (*) indicate a significance level of p < 0.05 compared to the expression levels in the HepG2 cell line.

Article Snippet: The NCI-H1975 (ATCC CRL-5908), NCI-H1650 (ATCC CRL-5883), NCI-H2228 (ATCC CRL-5935), and HepG2 (HB-8065) cell lines were obtained from the American Type Culture Collection (ATCC, USA).

Techniques: Expressing, Derivative Assay, RNA Sequencing, Quantitative RT-PCR

Journal: Food Science & Nutrition

Article Title: Fucoxanthin Ameliorates MASLD by Directly Targeting GRP78 to Restore ER Homeostasis and Activate AMPK Signaling

doi: 10.1002/fsn3.71813

Figure Lengend Snippet: Fucoxanthin directly targets and thermally stabilizes GRP78. (A) Schematic diagram of the drug affinity responsive target stability (DARTS) assay workflow used to identify fucoxanthin‐binding proteins. (B) Representative silver‐stained SDS‐PAGE gel of DARTS assay. HepG2 cell lysates were incubated with fucoxanthin (Fux, 100 μM) or DMSO for 1 h, followed by proteolysis with pronase (1:100 ratio) for 20 min. The black arrow indicates a protected protein band near 78 kDa, identified as GRP78 by MS. (C) Cellular thermal shift assay (CETSA). HepG2 lysates treated with Fux (100 μM) were heated at the indicated temperature gradient (37°C–60°C). Western blots show enhanced thermal stability of GRP78 in the presence of Fux. (D) Isothermal dose–response (ITDR) CETSA. Lysates were treated with increasing concentrations of Fux (10 −3 to 10 2 μM) at a fixed temperature (52°C), confirming dose‐dependent stabilization of GRP78.

Article Snippet: HepG2 human hepatoma cells were obtained from the American Type Culture Collection (ATCC, #HB‐8065) and cultured in DMEM supplemented with 10% (v/v) FBS and 1% penicillin/streptomycin in a humidified atmosphere containing 5% CO 2 at 37°C.

Techniques: Binding Assay, Staining, SDS Page, Incubation, Thermal Shift Assay, Western Blot

Journal: Food Science & Nutrition

Article Title: Fucoxanthin Ameliorates MASLD by Directly Targeting GRP78 to Restore ER Homeostasis and Activate AMPK Signaling

doi: 10.1002/fsn3.71813

Figure Lengend Snippet: Fucoxanthin mitigates palmitic acid (PA)‐induced ER stress in HepG2 cells. (A, B) Representative western blots and quantification of GRP78 (A) and p‐eIF2α/eIF2α ratio (B) in HepG2 cells treated with PA (100 μM) ± fucoxanthin (Fux) for 24 h. (C–F) Relative mRNA levels of ER stress‐related genes: HSPA5 (C), ATF6 (D), DDIT3 (E), and ERN1 (F). (G, H) Effect of Fux on thapsigargin (Tg)‐induced ER stress. Cells were treated with Tg (1 μM) ± Fux. (G) Western blots of GRP78 protein expression. (H) Intracellular triglyceride (TG) content normalized to total protein. Data are presented as mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: HepG2 human hepatoma cells were obtained from the American Type Culture Collection (ATCC, #HB‐8065) and cultured in DMEM supplemented with 10% (v/v) FBS and 1% penicillin/streptomycin in a humidified atmosphere containing 5% CO 2 at 37°C.

Techniques: Western Blot, Expressing

Journal: Food Science & Nutrition

Article Title: Fucoxanthin Ameliorates MASLD by Directly Targeting GRP78 to Restore ER Homeostasis and Activate AMPK Signaling

doi: 10.1002/fsn3.71813

Figure Lengend Snippet: GRP78 knockdown abolishes the lipid‐lowering effect of fucoxanthin. (A) Validation of GRP78 knockdown efficiency by western blot in HepG2 cells. GRP78 protein levels were reduced by approximately 66% in siGRP78‐transfected cells compared to siCtl group. Tubulin served as loading control. (B) Representative Oil Red O staining images (left) and quantification of lipid droplet area (right). GRP78 knockdown reversed the protective effect of Fux against PA‐induced lipid accumulation. (C) Quantification of intracellular TG levels. (D–G) Relative mRNA expression of lipid metabolism genes: Fatty acid oxidation genes PPARα (D) and CPT1 (E); lipogenic genes SREBP1C (F) and FASN (G). Data are presented as mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001, ns, not significant. Scale bar = 25 μm.

Article Snippet: HepG2 human hepatoma cells were obtained from the American Type Culture Collection (ATCC, #HB‐8065) and cultured in DMEM supplemented with 10% (v/v) FBS and 1% penicillin/streptomycin in a humidified atmosphere containing 5% CO 2 at 37°C.

Techniques: Knockdown, Biomarker Discovery, Western Blot, Transfection, Control, Staining, Expressing

Journal: PLOS One

Article Title: Unraveling the role of ChREBP in lung adenocarcinoma: Expression, regulatory networks, and potential functional impact

doi: 10.1371/journal.pone.0347907

Figure Lengend Snippet: (A) Expression levels of ChREBP ( MLXIPL ) in 66 human LUAD cell lines, derived from RNA-seq datasets available in the EMBL-EBI Expression Atlas, are presented in transcripts per million (TPM). (B-F) Expression of ChREBP in NCI-H1975, NCI-H1650, NCI-H2228 and HepG2 cells assessed using RT-qPCR assays. (B) Expression level of seven candidate HKGs. Cycle threshold (Ct) values are displayed. (C) Average expression stability values of seven HKGs determined by geNorm are shown. Expression levels of total ChREBP (D), ChREBP-α (E), and ChREBP-β (F) were normalized against the three most stable HKGs, RPS13, QARS, and RNA18S . Data represents the results of three independent experiments. Asterisks (*) indicate a significance level of p < 0.05 compared to the expression levels in the HepG2 cell line.

Article Snippet: The HepG2 cell line was maintained in Eagle’s Minimal Essential Medium (ATCC, USA) with 10% FBS (Gibco, USA).

Techniques: Expressing, Derivative Assay, RNA Sequencing, Quantitative RT-PCR