Panel A: Two rows of images showing primary tumor and organoid lines. The top row displays primary tumor images with H and E and immunostaining for Ar, PanCK, Vim, Insm1, and Syp. The bottom row shows corresponding organoid lines with the same staining. Panel B: A bar graph showing relative Sirt1 expression in organoid lines at baseline (shControl) and after silencing with two shRNA. The y-axis represents relative Sirt1 expression, and the x-axis shows the different conditions. The graph indicates a significant decrease in Sirt1 expression in the shSirt1 hashtag1 and shSirt1 hashtag 2 conditions compared to the shControl. Panel C: Bright-field and fluorescence images of NPp53 organoids under different conditions (Original, shControl, shSirt1 hashtag 1, shSirt1 hashtag 2). Images are shown at low and high power.
Establishment of an organoid model of NEPC from NPp53 mice. (A) Representative images of the parental tumor and the corresponding organoid lines derived from an NPp53 mouse and propagated in vitro. The images show H&E or immunostaining for the indicated markers. Scale bars represent 50 μm for the tumors and 20 μm for the organoids. (B) Real-time PCR showing relative expression of Sirt1 in the organoid line at baseline (shControl) and following silencing with two independent shRNA (shSirt1#1 or shSirt1#2). Data are normalized using Gapdh as internal control. The experiments were repeated three times with two biological replicates. (C) Representative bright-field and fluorescence images of NPp53 organoids as indicated. Scale bars represent 200 μm for low power and 50 μm for high power.