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Figure 8 Supplement: pTLK(S695) and pChk1(S345) western blots

Figure 8 Supplement: pTLK(S695) and pChk1(S345) western blots. 98 kd…. P-TLK(s695) ~ 86 kd. 64 kd…. Gem +LY. Vehicle. LY. Gem. 64 kd…. P-Chk1(s345) ~ 55 kd. 50 kd…. Figure 8 Supplement pChk1 (S296) western blot. G 5. L. L. G 6. G 4. G 5. G 3. G 3. G 7. G 1. L+G. L+G. V.

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Figure 8 Supplement: pTLK(S695) and pChk1(S345) western blots

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  1. Figure 8 Supplement: pTLK(S695) and pChk1(S345) western blots 98 kd… P-TLK(s695) ~ 86 kd 64 kd… Gem +LY Vehicle LY Gem 64 kd… P-Chk1(s345) ~ 55 kd 50 kd…

  2. Figure 8 Supplement pChk1 (S296) western blot G5 L L G6 G4 G5 G3 G3 G7 G1 L+G L+G V G2 G7 V L G1 G6 V G4 L+G V L+G V L G2 L+G V=Vehicle treated G=Gem alone treated L=LY2603618 alone treated L+G=LY+Gem combination treated

  3. Supplement to Figure 8: Representative Western Blot Images of graphed data Mice bearing Calu-6 xenografts were treated with vehicle, 150 mg/kg gemcitabine, 200 mg/kg LY2603618 or a simultaneous dose of gemcitabine and LY2603618. After 8 hours the tumors were removed, processed and the cell lysates equalized for total protein. See Materials and Methods. Protein lysates were separated and randomized by one operator using ePage™ 96 well 6% gels (Invitrogen) in the case of the pChk1(S296) data image, or by a second operator without randomizing samples, using 26 well 10% SDS-PAGE Criterion gels (Biorad) in the case of pTLK(S695) and pChk1(S345). Given the space for a multiple sample load in the 96 well pChk1(S296) gel/blot, lysates from gemcitabine treated animals were applied in duplicate (labeled as G1, G1 and G2, G2 etc.) and all samples were randomized throughout. Samples loaded on the 26 well gels/blots were not randomized nor duplicated. Molecular weight markers were run on both gel types to identify the specific protein band in the immunoblot.

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