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

Figure 1. Nf1 loss activates Fak1 and accelerates murine LUAD tumorigenesis.

Figure 1

  • A
    Schematic of Kras LSL‐G12D/+; p53 fl/fl (KP) mice intratracheally infected with pSECC lentiviruses containing sgNf1 or control sgTom. Mouse tumor burden was tracked by micro‐computed tomography (micro‐CT) for 5 months post‐infection, and lungs were finally harvested 21 weeks post‐infection. Tumors were dissected out for immunohistochemical (IHC) staining and generation of the tumor‐derived parental KP cell line.
  • B
    Quantification by micro‐CT of mean tumor volumes in KP mice derived from 50 randomly selected tumors at 8, 12, 16, and 20 weeks post‐infection with control sgTom, sgNf1.1, sgNf1.2, or sgNf1.3 (n = 18 mice, 21 mice, 21 mice, and 20 mice, respectively).
  • C
    Quantification by micro‐CT of total tumor volume per mouse in KP mice after infection with control sgTom, sgNf1.1, sgNf1.2, or sgNf1.3 at 8, 12, 16, and 20 weeks post‐infection (n = 18 mice, 21 mice, 21 mice, and 20 mice, respectively).
  • D
    Depictions and quantitation of total tumor burden (total tumor area/total lung area) in KP mice after infection with control sgTom, sgNf1.1, sgNf1.2, or sgNf1.3 at 21 weeks after infection (n = 18 mice, 21 mice, 21 mice, and 20 mice, respectively).
  • E
    Distribution of histological tumor grades in KP mice after infection with control sgTom, sgNf1.1, sgNf1.2, or sgNf1.3 (n = 50 tumors each) at 21 weeks after infection.
  • F
    Assessment of the mitotic index of tumor cells by phosphorylated‐histone H3 (pHH3)‐positive nuclei density in KP mice LUAD tumors at 21 weeks after infection with control sgTom, sgNf1.1, sgNf1.2, or sgNf1.3 (n = 50 tumors each).
  • G
    Nf1 mRNA expression in LUAD tumor sections 21 weeks after infection with control sgTom, sgNf1.1, sgNf1.2, or sgNf1.3 (n = 50 tumors each).
  • H
    Representative hematoxylin and eosin (H&E) and p‐Fak1 immunohistochemical (IHC) staining of LUAD tumor sections 21 weeks after infection with control sgTom (grade 1 depicted), sgNf1.1 (grade 3 depicted), sgNf1.2 (grade 3 depicted), or sgNf1.3 (grade 3 depicted) (n = 50 tumors each). H&E scale bars (low‐magnification top row = 100 μm, high‐magnification bottom row = 250 μm); p‐Fak1 IHC scale bars (low‐magnification top row = 250 μm, high‐magnification bottom row = 500 μm).
  • I
    Quantification of p‐Fak1 IHC signals in LUAD tumor sections 21 weeks after infection with control sgTom, sgNf1.1, sgNf1.2, or sgNf1.3 (n = 50 tumors each).
  • J
    Quantification of p‐Fak1 IHC signals in sgNf1.3 LUAD tumor sections analyzed by tumor grade (n = 50 tumors).
  • K
    Quantification of Psat1 mRNA expression in LUAD tumor sections 21 weeks after infection with control sgTom, sgNf1.1, sgNf1.2, or sgNf1.3 (n = 50 tumors each).
  • L
    Quantification of Psat1 mRNA expression in sgNf1.3 LUAD tumor sections analyzed by tumor grade (n = 50 tumors).
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.