Fig 1: shRNA‐mediated knockdown of Tyk2 in JW23.3 murine MPNST cells leads to decreased tumor growth in vivo. (A) Dissected tumors from subcutaneous injection of shTyk2‐infected JW23.3 cell lines and control shLacZ. (B) Quantification of subcutaneous tumor volume over time. (C) Quantification of the number of Cleaved Caspase‐3 positive cells from immunohistochemistry of harvested subcutaneous tumors of shTyk2‐infected JW23.3 cell lines and control shLacZ. (D) Representative images of Cleaved Caspase‐3 immunohistochemistry. (E) Tumor burden over time as measured by photon flux in mice injected intraventricularly. (F) Representative images of mice following intraventricular injection. (n = 5 for all groups; *P < 0.05, **P < 0.01, ***P < 0.001)
Fig 2: Tyrosine kinase 2 (TYK2) protein expression in malignant peripheral nerve sheath tumors (MPNSTs) and other sarcomas. TYK2 immunoreactivity was observed NF1‐associated and sporadic MPNSTs as well as other sarcomas. (A) Representative images of positive and negative staining for Plexiform Neurofibromas, NF1‐associated MPNSTs, sporadic MPNSTs, and Undifferentiated Pleomorphic Sarcoma (UPS), and fibrosarcoma. (B) Quantification of number of positive and negative cases observed for each type of tumor
Fig 3: Knockdown of Tyk2/TYK2 in JW23.3 murine MPNST and MPNST 724 cells affects expression of downstream targets. Western blot densitometry analysis on downstream targets of Tyk2/TYK2. Murine JW23.3 cell lines are depicted in blue/green, human MPNST 724 cell line are depicted in black/orange. (A and B) levels of TYK2 normalized to GAPDH (C and D) levels of p‐STAT1 normalized to total STAT1. (E and F) levels of p‐STAT3 normalized to total STAT3. (G and H) levels of Bcl‐2 normalized to GAPDH (I and J) levels of Cleaved Caspase‐3 normalized to Caspase 3 levels. (*P < 0.05, **P < 0.01, ***P < 0.001) All experiments were done in triplicate. Representative blots are shown
Fig 4: Loss of Tyk2/TYK2 in JW23.3 murine MPNST and MPNST 724 cells leads to increased cell death. shTyk2‐infected JW23.3 cell lines and control shLacZ: (A) Western blot densitometry analysis measuring TYK2 protein. (B) Relative Expression of Tyk2 mRNA compared to Gapdh. (C) An Incucyte cell proliferation assay measuring confluence over time. (D) An Incucyte death assay measuring TOTOTM‐3 iodide fluorescence as an indicator of death over time. TYK2 knockout MPNST 724 cells and scramble control: (E) Western blot densitometry analysis of TYK2 protein. (F) Relative Expression of TYK2 mRNA compared to GAPDH. (G) An Incucyte cell proliferation assay measuring confluence over time. (H) An Incucyte death assay measuring TOTOTM‐3 iodide fluorescence as an indicator of death over time. (*P < 0.05, **P < 0.01, ***P < 0.001)
Supplier Page from Abcam for Anti-TYK2 antibody