Extended Data Fig. 3: A3A drives widespread deaminase-independent CIN in PDA.
a, mRNA expression (2-ddCT) of A3A in MiaPaca-2 and Panc1 cells expressing A3AWTLV196 treated + /- Dox (n = 2 each). b, mRNA expression (2-ddCT) of A3A in MiaPaca-2 and Panc1 cells expressing inducible shRNA A3A and treated + /- Dox (n = 2 each). a, b, n=technical replicates. c, d, γH2AX staining and quantitation in MiaPaca-2 and Panc1 cells expressing A3AWTLV196 + /-Dox (n = 3 each) or inducible shRNA A3A + /- Dox (MiaPaca + /- Dox n = 3; Panc1 n = 3 (-Dox), n = 4 (+Dox); n=technical replicates. Data are represented as mean. e-g, % γH2AX positive cells within e-f, PKCY cells expressing indicated constructs of A3A (PKCY control (n = 5) versus A3AC161Y (n = 3) P = NS., control versus A3AE72A (n = 4) *P = 0.0159, control versus A3ATR (n = 3) *P = 0.0357, A3AWTLV196 + Dox versus -Dox (n = 3 each) P = NS. and within g, MiaPaca-2 +Dox versus -Dox (n = 2 each) and Panc1 +Dox versus -Dox (n = 2 each) cells expressing A3AWTLV196 or shRNA to A3A. n=biologically independent cells lines; e-f, Two-sided Mann–Whitney-U test. h-p, Giemsa staining with analysis of 35 or more metaphases of PKCY tumor cells expressing A3AWTLV196 + /- Dox (n = 4 each). q-y, Giemsa staining with analysis of 35 or more metaphases of PKCY tumor cells expressing A3AC161Y and control vector (n = 4 each). z-hh, Giemsa staining with analysis of 35 or more metaphases of PKCY tumor cells expressing A3AE72A and control vector (n = 3 each). ii-qq Giemsa staining with analysis of 35 or more metaphases of PKCY tumor cells expressing A3ATR and control vector (n = 2 each). rr, Representative images of Giemsa staining of primary A3A high and A3A low hPDX-derived cells. Scale bars = 100 μm. ss-zz, Giemsa staining with analysis of 35 or more metaphases of primary A3A high and A3A low hPDX-derived cell lines (n = 4 each); ss-zz: ***P < 0.0001; Chi-square test w/o Yate’s correction. Scale bars = 100 μm. h-zz, n=biologically independent cell lines. All insets at x80.