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. 2019 Apr 29;11(6):e9856. doi: 10.15252/emmm.201809856

Figure 2. Nf1 loss accelerates tumor growth in p53‐independent manner.

Figure 2

  • A
    Western blotting analysis of p53 expression in various p53/TP53‐WT cell lines incubated with the DNA‐intercalating agent doxorubicin (DOXO, 0.2 μg/ml for 6 h) to induce p53 stabilization (n = 3 biological replicates). Full experimental data provided in Appendix Fig S2A.
  • B, C
    Western blotting analysis confirming NF1 upregulation and p‐FAK1 downregulation in (B) DOXO‐treated PDKN1 cells and (C) DOXO‐treated PDKN2 cells transduced with PGK‐NF1 (n = 3 biological replicates). Full experimental data provided in Appendix Fig S2B and C.
  • D
    Western blotting analysis confirming NF1 downregulation and p‐FAK1 upregulation in DOXO‐treated SW1573 cells transduced with sgNf1.3 (n = 3 biological replicates). Full experimental data provided in Appendix Fig S2D.
  • E, F
    Western blotting analysis confirming Nf1 downregulation and p‐Fak1 upregulation in (E) DOXO‐treated LKR10 and (F) DOXO‐treated LKR13 clones transduced with sgNf1.3 (n = 3 biological replicates). Full experimental data provided in Appendix Fig S2E and F.
  • G
    FAK1 target gene expression in DOXO‐treated PDKN1 cells transduced with PGK‐control or PGK‐NF1 (n = 3 biological replicates).
  • H, I
    Subcutaneous tumor volumes of DOXO‐treated PDKN1 and PDKN2 cells transduced with PGK‐control or PGK‐NF1 (n = 18 each).
  • J–L
    Subcutaneous tumor volumes of (J) SW1573, (K) LKR10, and (L) LKR13 cells transfected with sgTom or sgNf1.3 (n = 18 each).
  • M
    Depictions and quantitation of Kras G12D/+; p53 +/+ (K‐only) autochthonous tumor burden (total tumor area/total lung area) in pSECC‐sgTom (n = 18 mice) or pSECC‐sgNf1.3 mice (n = 21 mice).
  • N
    Analysis of tumor grades in K‐only autochthonous tumors derived from pSECC‐sgTom (n = 18 mice) or pSECC‐sgNf1.3 mice (n = 21 mice).
  • O
    Quantification of Ki‐67‐positive nuclei per mm2 in K‐only autochthonous tumors derived from pSECC‐sgTom (n = 18 mice) or pSECC‐sgNf1.3 mice (n = 21 mice).
Data information: P‐values are reported in Appendix Table S3. In bar charts and line graphs, data presented as means with error bars representing standard deviations (SDs). For boxplots, whiskers indicate the minimum and maximum values, the upper and lower perimeters represent the first and third quartiles, the midline represents the median value, and the x symbol represents the mean.Source data are available online for this figure.