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The immunocomplexes were probed by western blotting with monoclonal anti-LKB1 (Sigma-Aldrich), SIRT1 (Abcam), SIRT6 (Abcam), or acetyl-lysine antibody (Cell Signaling). Statistical analysis. Data are presented as means ± S.D. Two-way analysis of variance for comparisons was performed for ...
Antibodies The following monoclonal antibodies used for staining and cell sorting were from BD Pharmingen: anti-CD4 (GK1.5 and RM4- 4), anti-CD8α (53-6.7), anti-TCRβ (H57-597), anti-CD44 (IM-7), anti-CD25 (3C7), anti-CD5 (53-7.3) and anti-CD2 (RM2-5). Anti- phospho-...
Par-3 was detected using a rabbit polyclonal anti- Par-3 antibody (Provided by Dr Tony Pawson, 1:200 dilution). LKB1 was detected by using a mouse monoclonal anti-LKB1 antibody (Abcam, 1:50 dilution), rabbit polyclonal anti-LKB1 antibody (Abcam, 1:50 dilution), mouse monoclonal anti- ...
[25]. For analysis of Lkb1 levels, cell lysates were diluted to a final concentration of 0.5 mg/mL and loaded in a microplate (Protein Simple #043-165), along with a monoclonal mouse anti-Lkb1 antibody (1:50; Invitrogen 206R24). Western blotting was conducted according to the instructions...
For analysis of Lkb1 levels, cell lysates were diluted to a final concentration of 0.5 mg/mL and loaded in a microplate (Protein Simple #043-165), along with a monoclonal mouse anti-Lkb1 antibody (1:50; Invitrogen 206R24). Western blotting was conducted according to the instructions for ...
clone sp6, rabbit monoclonal, #RM-9106-S, 1:500), Cleaved Caspase-3 (Cell Signaling, clone Asp175, rabbit polyclonal, #9661, 1:2000) Glutathione Peroxidase 4 (Abcam, rabbit monoclonal, ab125066, 1:2250), and γH2AX (Cell Signaling, Ser139 clone 20E3, rabbit monoclonal, #9718, 1:500...
The purified LKB1 farnesylated peptide (10 mg) was used by Dundee Cell Products to produce a mouse monoclonal antibody that strongly recognized this antigen. The hybridoma expressing the most sensitive immunoblotting antibody that recognized farnesylated LKB1 was selected and antibody purified accordingly ...
Monoclonal U2OS cell lines stably expressing GFP-WIPI3S, GFPWIPI3 or GFP-WIPI4 have been generated upon transient transfections using Lipofectamine 2,000 (Invitrogen; 11668-019) and G418 selection (0.6 mg ml−1) (refs 31, 36). Autophagy assay Cells were incubated in full (fed, F)...
G418 Sulfate (Life Technologies; 11811-098) was additionally added (0.6 mg ml À 1) to monoclonal U2OS cell lines stably expressing GFP-WIPI1, Figure 7 | WIPI4 interacts with ATG2, AMPK and ULK1. (a) U2OS cells transiently expressing GFP-tagged WIPI proteins and myc-ATG2A were ...