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. 2021 Jun 8;13(7):e14089. doi: 10.15252/emmm.202114089

Figure 5. Features of SHP099 tumor growth inhibition.

Figure 5

  • A
    Anti‐tumor activity of SHP099 against B16F10 tumors established s.c. in NOD scid (NSG) mice (n = 5/group); P values from two‐tailed Student’s t‐test; *P < 0.05, **P < 0.01. Error bars: ± SD.
  • B
    Extravasation of FITC‐dextran (green) in tumors established in NSG mice treated with SHP099; scale bar (50 µm). Representative images from 3 mice/group.
  • C
    Reduced CD31+ vascular area in SHP099‐treated tumors compared to controls (n = 5/group); ImageJ quantification. P values from two‐tailed Student’s t‐test; *P < 0.05. Error bars: ± SD.
  • D, E
    SHP099 reduces growth (D) and weight (E) of control (pLKO vector) and SHP2‐depleted (shSHP2) B16F10 tumors compared to untreated controls: pLKO untreated (n = 5), pLKO treated with SHP099 (n = 9), shSHP2 untreated (n = 5), and shSHP2 treated with SHP099 (n = 13). P values by analysis of variance with Dunnett’s multiple comparison test; ***P < 0.001.
  • F
    Persistent depletion of SHP2 in tumors induced by shSHP2‐B16F10 cells (n = 5) compared to control tumors induced by pLKO‐B16F10 (n = 5); tumors removed from mice 18 days after cell inoculation.
  • G
    SHP099 induces extravasation of FITC‐dextran (green) in tumors from inoculation of shSHP2‐B16F10 (representative image from 3 tumors); scale bar: 50 µm.
  • H
    Reduced CD31+ vascular area in SHP099‐treated tumors from s.c. inoculation of shSHP2‐B16F10 cells compared to controls from pLKO‐B16F10 cells (n = 5/group); ImageJ quantification. Error bars: ± SD; P values from two‐tailed Student’s t‐test; *P < 0.05.
  • I
    Schematic of experiment.
  • J, K
    After footpad inoculation of B16F10 cells, mice were treated orally with SHP099 (n = 20) or vehicle only (n = 20). Tumor size (J) was estimated from foot weight (g); size of popliteal lymph nodes (K) was measured (mm2) by ImageJ software; Student’s t‐test; **P < 0.01, ***P < 0.001.
  • L, M
    Proportion of mice in the SHP099 and control groups with macroscopic (L) and microscopic (M) evidence of metastases; Fisher’s exact test; ***P < 0.001.
  • N
    Representative popliteal lymph nodes from control and SHP099‐treated mice with microscopic evidence of metastases; H&E stain; scale bars 500 µm (low magnification) 20 µm (high magnification).