Extended Data Fig. 6: FN1 is a critical mediator of LPAR4-induced cancer stemness.
a, Gene Set Enrichment Analysis (GESA) for LPAR4-induced gene expression suggests that AKT signaling is upregulated in 79E + R4 cells in the absence of LPA. b, Immunoblot showing representative of three biological experiments for the protein levels of p-AKT-S473, AKT, p-GSK-3β-S9, p-CREB-S133, CREB, and vinculin in Colo-357+EV and Colo-357 + R4 cells treated with Ipatasertib of a range of doses for 2 h, or with 1 μM Ipatasertib in a time-course experiment. c, Relative mRNA level of ECM-related genes among LPAR4 gene signature in +R4 cells transfected with si-CREB as compared to cells transfected with si-CTRL. d, Graphs showing the cell viability of cells grown on 2D with serum free or with 10% charcoal stripped FBS containing media. Cell viability was assessed by the CellTiter-Glo assay, and all numbers were normalized to EV cells transfected with si-CTRL. e, Quantitative RT-PCR confirmation of FN1 knockdown by using siRNA in three pairs of +EV and +R4 cells as indicated. f, Representative histograms of three biological experiments showing MitoSOX signaling in +EV vs. +R4 cells treated with scrambled siRNA or si-FN1. All cells were cultured in serum free media for 48 h prior to MitoSOX staining. Data were presented as mean ± s.d. for n = 3 biological experiments (c–e). P-value was calculated using two tailed unpaired one sample t-test. Source numerical data are available in source data.
