g401 cell line  (ATCC)

Journal: Scientific Reports

Article Title: Flavokawain C suppresses nephroblastoma growth by inducing autophagy-mediated downregulation of FABP4 via AMPK/mTOR pathway

doi: 10.1038/s41598-026-42414-1

Figure Lengend Snippet: FKC inhibited the growth of G401 cells. ( A ) The cell viability of G401 treated with different concentrations of FKC (0, 1, 5, 10, and 15 μM) and ( B ) Stimulated with 10 μM FKC at different time point was detected by CCK-8 assay. ( C ) The clone formation assay was conducted after G401 cells were treated with different dose of FKC (0, 1, 5, 10, and 15 μM). ( D ) The proliferation of G401 cells treated with or without FKC was examined by EdU assay. Data are presented as mean ± SD of three independent experiments; * P < 0.05 , compared to the Control group. NS not significant.

Article Snippet: For the combined treatment of FKC and CQ (20 μM, Cat. HY-17589A, MCE), G401 cells were pre-treated with CQ for 6 h, followed by stimulation with FKC.

Techniques: CCK-8 Assay, Tube Formation Assay, EdU Assay, Control

Journal: Scientific Reports

Article Title: Flavokawain C suppresses nephroblastoma growth by inducing autophagy-mediated downregulation of FABP4 via AMPK/mTOR pathway

doi: 10.1038/s41598-026-42414-1

Figure Lengend Snippet: FKC restrained the migration and invasion of G401 cells. ( A ) G401 cells were treated with different concentrations of FKC (0, 1, 5, and 10 μM), and the expression of epithelial-mesenchymal transition (EMT) markers was detected by western blot analysis. ( B ) G401 cells were stimulated with 10 μM FKC, and wound closure was detected by wound healing assay. ( C , D ) The inhibitory effect of FKC on the migration and invasion of G401 cells was detected by transwell assay. The bar graphs show the relative protein expression levels normalized to β-tubulin. Data are presented as mean ± SD of three independent experiments; * P < 0.05 , compared to the Control group.

Article Snippet: For the combined treatment of FKC and CQ (20 μM, Cat. HY-17589A, MCE), G401 cells were pre-treated with CQ for 6 h, followed by stimulation with FKC.

Techniques: Migration, Expressing, Western Blot, Wound Healing Assay, Transwell Assay, Control

Journal: Scientific Reports

Article Title: Flavokawain C suppresses nephroblastoma growth by inducing autophagy-mediated downregulation of FABP4 via AMPK/mTOR pathway

doi: 10.1038/s41598-026-42414-1

Figure Lengend Snippet: FKC suppressed growth and metastasis of G401 cells by decreasing FABP4 expression. The expression of FABP4 in nephroblastoma tissues and normal tissues was analyzed by western blot ( A ). G401 cells were treated with different concentrations of FKC (0, 1, 5, and 10 μM), and the expression of FABP4 was examined by Western blotting ( B ). G401 cells infected with oe-NC and oe-FABP4 lentivirus, then stimulated with or without FKC (10 μM). The efficiency of overexpression lentivirus was identified by Western blotting ( C ). The cell viability was assessed by CCK8 ( D ). Expression of E-cadherin, N-cadherin and Vimentin were assessed by Western blotting ( E ). The migration and invasion of G401 cells was were evaluated by transwell ( F , G ) and wound healing assays ( H ). The bar graphs show the relative protein expression levels normalized to β-tubulin. Data are presented as mean ± SD of three independent experiments; * P < 0.05 . NS not significant.

Article Snippet: For the combined treatment of FKC and CQ (20 μM, Cat. HY-17589A, MCE), G401 cells were pre-treated with CQ for 6 h, followed by stimulation with FKC.

Techniques: Expressing, Western Blot, Infection, Over Expression, Migration

Journal: Scientific Reports

Article Title: Flavokawain C suppresses nephroblastoma growth by inducing autophagy-mediated downregulation of FABP4 via AMPK/mTOR pathway

doi: 10.1038/s41598-026-42414-1

Figure Lengend Snippet: FKC induced autophagy in G401 cells. G401 cells were treated with 10 μM FKC for different time (0, 4, 8, 12, 18, and 24 h), and the expression of LC3 was detected by western blot ( A ). G401 cells were treated with 10 μM FKC and/or 5 µM CQ for 12 h, and the expressions of LC3 and P62 were quantified by western blot ( B ). The autophagy flux after different treatments was observed by mRFP-GFP-LC3 adenovirus double label assay ( C ). The bar graphs show the relative protein expression levels normalized to β-tubulin. Data are presented as mean ± SD of three independent experiments; * P < 0.05 . NS not significant.

Article Snippet: For the combined treatment of FKC and CQ (20 μM, Cat. HY-17589A, MCE), G401 cells were pre-treated with CQ for 6 h, followed by stimulation with FKC.

Techniques: Expressing, Western Blot

Journal: Scientific Reports

Article Title: Flavokawain C suppresses nephroblastoma growth by inducing autophagy-mediated downregulation of FABP4 via AMPK/mTOR pathway

doi: 10.1038/s41598-026-42414-1

Figure Lengend Snippet: Inhibition of autophagy attenuated the inhibitory effect of FKC on FABP4 expression. G401 cells transfected with sh-Atg5 or sh-NC were treated with FKC. The expression of autophagy-related proteins was detected by western blot ( A ). The expressions of FABP4 after sh-Atg5 transfection were evaluated by western blot analysis ( B ). The interaction between p62 and FABP4 in FKC -treated G401 was evaluated by Co-IP assay ( C ). The bar graphs show the relative protein expression levels normalized to β-tubulin. Data are presented as mean ± SD of three independent experiments; * P < 0.05.

Article Snippet: For the combined treatment of FKC and CQ (20 μM, Cat. HY-17589A, MCE), G401 cells were pre-treated with CQ for 6 h, followed by stimulation with FKC.

Techniques: Inhibition, Expressing, Transfection, Western Blot, Co-Immunoprecipitation Assay

Journal: Scientific Reports

Article Title: Flavokawain C suppresses nephroblastoma growth by inducing autophagy-mediated downregulation of FABP4 via AMPK/mTOR pathway

doi: 10.1038/s41598-026-42414-1

Figure Lengend Snippet: FKC induced autophagy in G401 cells via the AMPK/mTOR signaling pathway. G401 cells were treated with or without FKC for 30 min, and the expressions of proteins in AMPK/mTOR pathway were detected by western blot ( A ). G401 cells were treated with FKC and/or BAY-3827 for 24 h, followed by western blot analysis to detect the expressions of autophagic markers and FABP4 ( B ). The bar graphs show the relative protein expression levels normalized to β-tubulin. Data are presented as mean ± SD of three independent experiments; * P < 0.05.

Article Snippet: For the combined treatment of FKC and CQ (20 μM, Cat. HY-17589A, MCE), G401 cells were pre-treated with CQ for 6 h, followed by stimulation with FKC.

Techniques: Western Blot, Expressing

Journal: Scientific Reports

Article Title: Flavokawain C suppresses nephroblastoma growth by inducing autophagy-mediated downregulation of FABP4 via AMPK/mTOR pathway

doi: 10.1038/s41598-026-42414-1

Figure Lengend Snippet: FKC inhibited the growth and metastasis of nephroblastoma in vivo. Nude mice were subcutaneously injected with normal and oeNC or oeFABP4 infected G401 cells. One week after cell injection, mice in the treatment group were intraperitoneally administered 3 mg/kg FKC thrice a week for 4 weeks; control mice received the same volume of normal saline. Images of tumor masses from each group ( A ). The value of tumor volume and weight was quantified in each group ( B – C ). Tumor tissue sections stained with hematoxylin–eosin, the expression of Ki67, FABP4 and Vimentin by immunohistochemistry ( D ). Western blot was performed to detect the expression of FABP4, E-Cadherin and Vimentin in animal samples ( E ). The bar graphs show the relative protein expression levels normalized to β-tubulin. All data are presented as mean ± SD. * P < 0.05, NS not significant.

Article Snippet: For the combined treatment of FKC and CQ (20 μM, Cat. HY-17589A, MCE), G401 cells were pre-treated with CQ for 6 h, followed by stimulation with FKC.

Techniques: In Vivo, Injection, Infection, Control, Saline, Staining, Expressing, Immunohistochemistry, Western Blot

Journal: Science Advances

Article Title: IRS4 is a PI3K-activating cancer dependency up-regulated through DNA rearrangements or epigenetic mechanisms in multiple solid tumors

doi: 10.1126/sciadv.aeb3503

Figure Lengend Snippet: ( A ) Violin plots of gene dependency for n = 1077 cell lines (DepMap) for genes targeted by FDA-approved targeted therapies (blue) or chemotherapies (red). Genes inhibited by multiprotein-targeted inhibitors were excluded. Individual cell lines (points) are shown when ≤−1.0 (not shown when median gene dependency ≤−0.5). ( B ) Box plot of median dependencies for 17,453 genes, separated into FDA-approved classes (A) or other genes. P values, two-sided Wilcoxon rank-sum test. n.s., not significant. ( C ) Box plot of z -scores for the GPD ξ value for each gene’s dependency distribution as a measure of selective dependency. Genes above the top quartile of the targeted therapy group (1.98%; n = 346) were selected for further analysis. ( D ) Workflow to identify pHTI genes using selective dependency (C), human and mouse genetics, and normal tissue (GTEx) expression. ( E ) Box plot of IRS4 dependency scores by cancer type. ( F ) Correlation between IRS4 mRNA (CCLE RNA-seq) and dependency scores. r , Pearson correlation. ( G ) Western blot of indicated cell lines. ( H ) Western blot 3 days posttransduction with control or gene-targeted lentiviral sgRNAs at 10 MOI (for combination, each sgRNA at 5 MOI). Puromycin began 1 day posttransduction. ( I to L ) Viability (CellTiter-Glo) after lentiviral transduction in IRS4-absent (I and K) and IRS4-expressing (J and L) cell lines. Cells were transduced at 10 MOI (TC797, PER-624, G401, and HCC2429) or 20 MOI (all others), puromycin selected for 1 or 2 days, and plated with six technical replicates per time point (or three for PER-704). Eight to 14 hours after plating at day 0, measurement was performed for normalization. For TC797, PER-624, G401, and HCC2429, a combination of two IRS4 or PLK1 sgRNAs was given; puromycin was maintained during days 0 to 3 for each of these, except G401. For other cell lines, sgRNAs were administered individually without puromycin maintenance postplating. Points, mean values; errors bars, standard deviation.

Article Snippet: G401, RPMI2650, BT474, and SKBR3 cells were obtained from American Type Culture Collection.

Techniques: Expressing, RNA Sequencing, Western Blot, Control, Transduction, Standard Deviation

Journal: bioRxiv

Article Title: Leptin Acts as a Peripheral Tropic Signal to Tune Steroidogenesis

doi: 10.64898/2026.04.13.718290

Figure Lengend Snippet: (A) Fold-change plot of differentially expressed genes (DEGs) is shown with log2 fold change values and p-value color coding from RNA-seq analysis comparing early versus late L6 nymph in the PG. (B) Representative confocal images and quantification (C) of HSL protein levels in mouse Leydig cells from young (5 weeks), adult (6 months), and tumor-bearing mice. Scale bar: 20 μm. (D) qPCR analysis of relative leptin mRNA levels in HEK293T cells compared to G401 cells, showing significant upregulation of leptin signaling in the tumor context. (E) Bodipy staining (green) of steroidogenic tissues in control (6 months) and tumor-bearing mice (4 months). (F) Quantitative analysis of LD area in Leydig cells based on image E. (G) A schematic model illustrating the mechanism across D. melanogaster , B. germanica , and M. musculus . The leptin/Upd-JAK/STAT-HSL axis acts as a universal spatiotemporal switch, coordinating lipid mobilization to transition steroidogenic cells from a quiescent (LD-poor) to an activated (LD-rich) state during development or tumor-induced stress. Data are presented as mean ± s.e.m.. Statistical significance was determined by one-way ANOVA followed by multiple comparisons. **p < 0.01, ****p < 0.0001.

Article Snippet: G401 cells were cultured in McCoy’s 5A medium (ATCC, 30-2007TM) supplemented with 10% FBS.

Techniques: RNA Sequencing, Staining, Control