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. 2021 May 10;6(9):e140382. doi: 10.1172/jci.insight.140382

Figure 7. AMACR promotes CRC cell differentiation in the context of LIN28B overexpression.

Figure 7

(A) qPCR for intestinal differentiation markers in Caco-2 control/AMACR KD Caco-2 cells (n = 3). (B) ALP activity assay in Caco-2 control/AMACR KD cells (n = 3). (C) Upper: WB analysis of AMACR and CK20 expression in Caco-2 cells with control or si-AMACR at the confluence time point. Lower: densitometry; the value for CK20 or CDX2 at –2 days for the sh-control samples was designated as 1. (D) Left: representative image for dome formation in Caco-2 cell at postconfluence day 3. Scale bars: 500 μm. Right: The graph indicates the number of domes. A dome was defined as greater than 100 μm diameter. (n = 5) (E) Representative IHC staining images of human CRC TMAs. (F) The ATP assay in Caco-2 with control and LIN28B KD cells (n = 4). (G) The ATP assay in Caco-2 with control and AMACR KD cells (n = 4). (H) Butyric acid measurement in Caco-2 with control and AMACR KD cells (n = 6). (I) ALP activity assay in Caco-2 control/AMACR KD cells with or without 1 mM sodium butyrate (Na-B) (n = 3). Data are presented as mean ± SEM. Unpaired, 2-tailed Student’s t test (F) and 1-way ANOVA followed by Dunnett’s multiple-comparison test as post hoc analysis (A–D and G–I) were performed. *P < 0.05, **P < 0.01.