Journal: bioRxiv
Article Title: Hyper-responsiveness of cancer stem cells to microenvironmental cues controls metastasis and therapy response through YAP/TAZ/TEAD
doi: 10.1101/2025.03.13.643008
Figure Lengend Snippet: a. Flow cytometry for LPAR1 expression in LM2 CSCs (mCherry pos ) vs nonCSCs (mCherry neg ). b. Quantitation of data in (a). Mean +/− SD, n=3 c. Quantitation of YAP localization in LM2 CSCs vs nonCSCs in unsorted cultures with and without treatment with LPA assessed by immunofluorescent staining (n=524 and 400 for nonCSCs −/+ LPA; n=55 and 80 for CSCs −/+ LPA). Kruskal-Wallis test for CSCs v nonCSCs within treatment group. d. Top 10 enriched transcription factor binding motifs in differentially accessible chromatin regions in sorted LM2 CSCs vs nonCSCs around transcriptional start sites (TSS) or enhancer regions for 2 independent ATAC-Seq experiments. N* = NANOG. (Run 2 only had 9 enriched motifs that were statistically significant.) e. Genes with TEAD binding sites in their enhancers that are preferentially accessible in CSCs. Table shows overlap results of two independent runs (annotated genes only). f. RTQ-PCR for effect of TEAD inhibitor GNE7883 treatment on expression of KLF5 and ID1 in LM2 cells. CTGF is a canonical YAP target as positive control. Results are mean +/− SD for n=3, normalized to vehicle control. Students t-test. g. Flow cytometry for effect of 1D1 or KLF5 knockdown on % CSCs in LM2 cultures. Results are mean +/− SD for n=3, Dunnets multiple comparisons test vs control (CON). h. Schematic for mechanisms underlying hypersensitivity of CSCs to microenvironmental signals.
Article Snippet: Antibodies used were: LPAR1 (Alomone Labs LTR, Cat# ALR-031).
Techniques: Flow Cytometry, Expressing, Quantitation Assay, Staining, Binding Assay, Positive Control, Control, Knockdown