Figure 4 from Single-Cell Atlas of Lineage States, Tumor Microenvironment, and Subtype-Specific Expression Programs in Gastric Cancer

Abstract

<p>scRNA-seq enables identification of distinct gastric cancer fibroblast subtypes and <i>INHBA</i>–<i>FAP</i> axis as a CAF regulator. <b>A,</b> Bubble plots demonstrating stage-dependent increases in the proportion of fibroblast cells with STF3 being the dominant subcluster. The size of the circle represents the proportion of cells expressing subcluster-specific genes. <b>B,</b> Bubble plots showing fibroblast subclusters (STF1–3) expressing distinct CAF canonical markers (<i>FAP, CSPG4, ACTA2</i>, and <i>TAGLN</i>). The size of the circle represents the proportion of cells expressing different genes. <b>C,</b> Violin plot showing the expression of <i>INHBA</i> in STF2 and STF3 fibroblast clusters with negligible expression in the STF1 fibroblast cluster. <b>D,</b> Fibroblast ROIs captured by DSP analysis based on immunofluorescence staining for Pan-CK (epithelial, green), CD138 (plasma, pink), SMA (fibroblast, cyan), and DAPI (blue). The circular ROI is 300 μm in diameter. <b>E,</b> Bee swarm plot showing differential expression of <i>FAP</i> in fibroblast ROIs of normal and tumor samples by DSP (<i>n</i> = 13). <b>F,</b> Bee swarm plot showing differential expression of <i>INHBA</i> in fibroblast ROIs of normal and tumor samples by DSP (<i>n</i> = 13). <b>G,</b> Pearson correlation graph demonstrating strong positive correlations between <i>INHBA</i> and <i>FAP</i> gene expression in fibroblast ROIs using DSP. <b>H,</b> Bar graph showing significant expression of <i>FAP</i> and <i>INHBA</i> genes in flow-sorted tumor fibroblasts compared with matched normal fibroblasts (<i>n</i> = 10 each). <b>I,</b> Bar graph showing significant reduction in <i>FAP</i> gene expression after siRNA-mediated <i>INHBA</i> knockdown in tumor fibroblast lines. Two independent siRNAs were used. <b>J,</b> Bar graph showing significant increases in <i>FAP</i> gene expression in two normal fibroblast lines after treatment with recombinant INHBA (rINHBA) for 48 and 96 hours, respectively. <b>K,</b> Bubble plot depicting stage-dependent increases of <i>FAP</i><sup>+</sup> and <i>INHBA</i><sup>+</sup> cells in fibroblast cluster STF3 (<i>P</i> = 0.041). The circle sizes represent the relative proportion of cells expressing these genes. <i>P</i> values were computed using Kendall's τ method. <b>L,</b> Kaplan–Meier survival curves of TCGA-STAD data showing significant differences in overall survival between <i>INHBA</i>-high and <i>INHBA</i>-low samples. <i>P</i> values were computed using log-rank tests.</p>

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