g401  (ATCC)

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