Use of in Situ Isopropoxide Protection in the Metal−Halogen Exchange of Arylboronates

Isopropoxide protection of arylboronates allowed their use in metal−halogen exchange reactions. The isopropoxide-protected borate species were obtained from a boronate or in situ from dibromoarenes. meta- and para-dibromoarenes were converted via these intermediates into functionalized arylboronates in a one-pot manner

Use of in Situ Isopropoxide Protection in the Metal−Halogen Exchange of Arylboronates

Isopropoxide protection of arylboronates allowed their use in metal−halogen exchange reactions. The isopropoxide-protected borate species were obtained from a boronate or in situ from dibromoarenes. meta- and para-dibromoarenes were converted via these intermediates into functionalized arylboronates in a one-pot manner

Supplementary Figure 6 from GSK-3α Promotes Oncogenic KRAS Function in Pancreatic Cancer via TAK1–TAB Stabilization and Regulation of Noncanonical NF-κB

PDF file - 118K, GSK-3 inhibition leads to a decrease in TAK1 protein Panc-1 and MiaPaCa-2 cells were treated with GSK-3 inhibitor, AR-A014418 or vehicle control, DMSO in the presence or absence of MG132 for 24 hours. Whole cell extracts were immunoblotted for indicated antibodies.</p

Supplementary Figure 5 from GSK-3α Promotes Oncogenic KRAS Function in Pancreatic Cancer via TAK1–TAB Stabilization and Regulation of Noncanonical NF-κB

PDF file - 63K, GSK-3 inhibition inhibits TAK1 mediated signaling. Panc-1 cells were treated with GSK-3 inhibitor, AR-A014418 (30M) for 24 hours and whole cell lysates immunoblotted for indicated antibodies.</p

Supplementary Figure 3 from GSK-3α Promotes Oncogenic KRAS Function in Pancreatic Cancer via TAK1–TAB Stabilization and Regulation of Noncanonical NF-κB

PDF file - 113K, TAK1 inhibition decreases cell proliferation in Kras+ cells. Indicated cell lines were seeded in E-plate-16 (Roche) at 3000 cells/well/100μl. 24hours after plating, cells were treated with TAK1 inhibitor 5Z-7-oxozaenol (0.625μM), or vehicle control, DMSO. Cell impedance was measured every 2 hours for the entire course of experiment using RTCA.</p

Supplementary Methods from GSK-3α Promotes Oncogenic KRAS Function in Pancreatic Cancer via TAK1–TAB Stabilization and Regulation of Noncanonical NF-κB

PDF file - 72K</p

Supplementary Figure 7 from GSK-3α Promotes Oncogenic KRAS Function in Pancreatic Cancer via TAK1–TAB Stabilization and Regulation of Noncanonical NF-κB

PDF file - 85K, NF-κB2 depletion leads to increase in PARP cleavage. Indicated cell lines were transiently transfected with specified siRNAs, and 72 hours later whole cell extracts immunoblotted for cleaved PARP.</p

Supplementary Figure 2 from GSK-3α Promotes Oncogenic KRAS Function in Pancreatic Cancer via TAK1–TAB Stabilization and Regulation of Noncanonical NF-κB

PDF file - 181K, Left: Kras regulates TAK1 levels. HKRAS cells were transiently transfected with non-targeting or Kras siRNA for 72 hours. TAK1 was immunoprecitated from whole cell extracts and immunoblotted for specified antibodies. Right: TAK1 inhibition induces apoptosis. MiaPaCa-2 cells were treated with the TAK1 inhibitor, 5Z-7-oxozaenol at indicated concentrations for 24 hours. Whole cells extracts were immunoblotted with specified antibodies.</p

Supplementary Figure 1 from GSK-3α Promotes Oncogenic KRAS Function in Pancreatic Cancer via TAK1–TAB Stabilization and Regulation of Noncanonical NF-κB

PDF file - 308K, Left: AR-A014418 inhibits GSK-3 activity in a dose dependent mannerPanc-1 cells were treated with increasing concentrations of GSK-3 inhibitor, AR-A014418, like in Fig.1A. Whole cells extracts were prepared and immunoblotted for specified antibodies. p-Glycogen synthase is a downstream substrate of GSK-3 and thus an indicator of its kinase activity. Right: GSK-3 regulates apoptosis in MiaPaCa-2 cells. . Panc-1 and MiaPaCa-2 cells were transiently transfected with indicated siRNAs for 72 hours. Whole cell extracts were immunoblotted for specified antibodies.</p

Supplementary Figure 8 from GSK-3α Promotes Oncogenic KRAS Function in Pancreatic Cancer via TAK1–TAB Stabilization and Regulation of Noncanonical NF-κB

PDF file - 1842K, GSK-3 inhibition leads to a decrease in TAK1 levels and p100-p52 processing in a statistically significant manner. All data obtained with independent experiments with Panc-1 and MiaPaCa-2, treated with GSK-3 inhibitor for 24 hours (right panel) or transiently transfected with indicated siRNA's for 72 hours (left panel) were quantified by densitometric analyses and is expressed as mean�SD. Statistical evaluation of the differences were calculated using students two-tailed t test and P value < 0.05 was considered significant.</p