Fig. 3.

Loss of p53 and Tsc1 promote acinar cell transformation. (a) Kaplan–Meier survival analysis shows p53 ^-/-; Tsc1 ^-/- mice survival time (median survival: 101 days; n = 7) which is not different from that of Tsc1 ^-/- mice (median survival: 133 days; n = 5); (b) Representative H&E-stained sections of p53 ^-/-; Tsc1 ^-/+ and p53 ^-/-; Tsc1 ^-/- pancreata show pancreatic acinar cells with nuclear abnormalities (upper panel) and nodular hyperplasia (lower panel); scale bar: 50 μm (upper panel) and 200 μm (lower panel); (c) Representative H&E-stained sections and IHC pictures show the histology of one ACC-like tumor with a high proliferative index (p-HH3); (d-g) Representative IHC pictures show distinct in vivo activation of mTOR signaling (p-mTOR^Ser2448 and p-S6^Ser235/236), but inactivation of Erk (p-Erk1/2^{Thr202/Tyr204}) and Akt (p-Akt^Ser473) in tumor cells (and atypical acinar cells) of pancreatic tissues from p53 ^-/-; Tsc1 ^-/- mice, scale bar: 50 μm; (h) Phase contrast images of isolated cell lines from p53 ^-/- ; Tsc1 ^-/+ (911 and 961) and p53 ^-/- ; Tsc1 ^-/- (946 and 946 F) mice show fibroblast-like (961, 911 and 946 F) and epithelial (946) morphologies, scale bar: 50 μm; (i) Western-blot analysis demonstrates expression of Tsc1, E-Cadherin, Vimentin and p53 in p53 ^-/- ; Tsc1 ^-/+ and p53 ^-/- ; Tsc1 ^-/- cells; previously described cystic cells lines isolated from P48 ^Cre/+ ; Pten ^-/- ; Tsc1 ^-/+ mice were used as used as external controls [18]; (j) Western-blot analysis demonstrates phosphorylation levels of p-Akt^Ser473, p-Erk1/2^{Thr202/Tyr204} and p-mTOR^Ser2448 in p53^-/- ; Tsc1 ^-/+ and p53 ^-/- ; Tsc1 ^-/- cells after fetal bovine serum (FBS) treatment for 2 h; one of the three independent experiments is shown