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Abl and Arg Kinase Activity Regulate Melanoma Invasion and Proliferation

This study explores the role of Abl and Arg kinases in regulating melanoma invasion and proliferation. The results show that targeting Abl and Arg kinases can inhibit melanoma progression.

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Abl and Arg Kinase Activity Regulate Melanoma Invasion and Proliferation

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  1. Supplementary Figure S1 A Panel 1 Panel 2 melanocytes melanocytes SK-Mel-2 UACC-257 UACC-903 Mel-1617BR Malme-3M SK-Mel-5 SK-Mel-28 WM3248 Mel-1617 435s WM3211 WM164 451-Lu WM793 1205-Lu WM35 WM1232 LOX IMVI UACC-62 435s M14 WM9 WM3248 WM278 WM239 NIH3T3 NIH3T3 A375 sbcl2 Abl Kinase Assay Arg Kinase Assay pCrkL 35kD Abl 130kD Arg 130kD b-actin 40kD 64kD pAkt pSrc (Y416) 55kD c-Kit 180kD EGFR 100kD IGF1-R 180kD PDGFR-a 180kD PDGFR-b B pCrkL (Relative to Actin) Relative Abl+Arg Activity pSrc (Relative to Actin) Relative Abl+Arg Activity pSrc (Relative to Actin) C D E Matrigel Invasion Matrigel Invasion 3H-Thymidine * WM3248 * Relative Invasion * * * * ** ** Relative Proliferation * ** ** ** ** ** * * ** ** * *** * ** *** *** *** ** *** shRNA: Scr Abl/Arg Abl 130kD Arg 130kD β-actin 40kD siRNA siRNA LOX-IVMI UACC-903 siRNA: Scr Abl Arg Scr Abl Arg Abl 130kD Arg 130kD β-actin 40kD

  2. Supplementary Figure S2 A Abl BRAF Abl/BRAF/DAPI 20μm 20μm 20μm B Abl BRAF Abl/BRAF/DAPI 20μm 20μm 20μm C Abl1/Abl2-/- MEFs Abl BRAF Abl/BRAF/DAPI 20μm 20μm 20μm

  3. Supplementary Figure S3 WM3248 WM3248 A B antagonism additivity synergism

  4. Supplementary Figure S4 siRNA#2 shRNA A B WM3248 WM3248 UACC-903 Scr Abl/Arg-shRNA Scr Abl Arg Scr Abl Arg 40kD 40kD p-FRA-1 p-FRA-1 40kD 40kD FRA-1 FRA-1 25kD 25kD TWIST1 TWIST1 ZEB1 ZEB1 180kD 180kD 180kD 180kD ZEB2 ZEB2 40kD 40kD pERK pERK 40kD ERK 40kD ERK Abl Abl 130kD 130kD Arg 130kD Arg 130kD β-actin 40kD 40kD β-actin Plasmid: Scr Scr AA-1 AA-1 AA-2 AA-3 IPTG: - + - + + + Abl 130kD Arg 130kD 40kD β-actin C WM3248 Vector Abl-PP Arg-PP PLX (2h) : - + - + - + 25kD TWIST1 40kD p-FRA-1 40kD FRA-1 ZEB1 180kD 180kD ZEB2 pERK 40kD ERK 40kD Abl 130kD Arg 130kD β-actin 40kD

  5. Supplementary Figure S5 A WM3248 vehicle MK (10μM) nil (1μM) MK (10μM) +nil (1μM) vehicle MK (5μM) B UACC-903 nil (5μM) MK (5μM) +nil (5μM)

  6. Supplementary Figure S6 PLX (24h) A B WM3248 UACC-903 siRNA: Scr Abl Arg Scr Abl Arg 100kD 100kD BRAF BRAF b-actin 40kD b-actin 40kD WM3248 vehicle Abl-PP Abl/Arg-PP Abl-PP Abl/Arg-PP Vector Arg-PP Arg-PP Vector

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