Fig 2: Cdkn2b inactivation is indispensable for Rb phosphorylation of ductal cells. (a) Immunohistochemistry staining for total Rb and Rb phosphorylation at S780 was conducted on fresh adjacent sections from pancreases infected with the pTomo empty vector, KRAS-shp53-EGFP, KRAS-shp53-EGFP-shCdkn2a, KRAS-shp53-EGFP-shCdkn2b or KRAS-shP53-EGFP-shCdkn2a/b lentiviral particles. Scale bars: 20 µm. (b) The ratio of Rb phosphorylation-positive cell to EGFP-positive cells in a. (c) Immunohistochemistry staining for Rb1 phosphorylation in mouse pancreatic cancer. Scale bars: 40 µm. The quantification value is presented as the mean±s.e.m. ***P<0.001.

Fig 3: Poldip2 deletion does not affect Rb expression/phosphorylation.(A) Immunoblotting was performed using lysates from Poldip2+/+ and Poldip2-/- MEFs in passages 2, 4 and 5. The blots were probed with antibodies against Rb, pRb S780, pRb S807/811, pRb T821, Cdk2, and Cdk4. Densitometry was performed and corrected to ß-actin run in parallel (shown in Figure 7A) (B–G). (H) Phosphorylated proteins from Poldip2+/+ and Poldip2-/- cells in passages 2, 4 and 5 were purified with a phosphoprotein binding column. Phosphorylated Rb was measured by immunoblot and normalized to pAkt levels (I). Error bars represent mean ± SEM of 3–4 independent experiments. * P<0.05 comparing Poldip2+/+ with Poldip2-/-.

Fig 4: Expression of autophagic protein at different time points of wound healing. (A) Western blot analysis was used to detect related protein expression; (B) expression of EIF2AK2 protein; (C) expression of RB1 protein. *P<0.05. EIF2AK2, eukaryotic translation initiation factor 2 alpha kinase 2; RB1, RB transcriptional corepressor 1.

Fig 5: Single-sample gene set enrichment analysis immune cell infiltration analysis during early wound healing. (A) Box plot of the level of immune cell infiltration in the wound on day 0 and day 7; (B) box plot of the level of immune cell infiltration in the wound on day 0 and day 14; (C) correlation analysis heatmap of the expression level of EIF2AK2 and RB1 on immune cell infiltration in the wound on day 0 and day 7; (D) correlation analysis heatmap of the expression level of EIF2AK2 and RB1 and immune cell in the wound on day 0 and day 14. *P<0.05, **P<0.01, ***P<0.001. EIF2AK2, eukaryotic translation initiation factor 2 alpha kinase 2; RB1, RB transcriptional corepressor 1; ns, not significant.