Intracellular extracts of Jm-cadA and Jm-cadA-blc-Tfu showed a clear band corresponding to CadA-FLAG (82 kDa) (Figure 1, lanes 1 and 3).
Control cells showed no detectable rtTA expression in the absence or presence of doxycycline (Figure 1 lanes 1).
These also were not detected (Figure 1, Lanes D G), using the primers listed in Table 1.
We found that both CPZ and QC inhibited TBSV accumulation by ∼70 and 50%, respectively, when applied prior to electroporation (Fig. 1, lanes 9 and 13).
In each case, the immunoreactive bands were abolished after adsorption with the immunizing peptides (fig. 1, lanes 2, 4, 6, 8, 10, 12, 14).
In the absence of UTP, the polymerase is forced to pause at the first non-template T stretch (135 nt = T3) (Figure 1, lanes 1 and 2).
They appeared as prominent bands of 53 kD for CB1, (fig. 1, lane 1), 50 kD for CB2 (fig. 1, lane 3), 35 kD for MAGL (fig. 1, lane 5), 120 and 73 kD for DAGLα and DAGLβ respectively (fig. 1, lanes 7 and 9), and 46 and 63 kD for NAPE-PLD and FAAH respectively (fig. 1, lanes 11 and 13).
Immunoblotting showed endogenous PP6R1 and PP6c co-immunoprecipitated with endogenous DNA-PKcs from the proficient M059K cells using specific anti-DNA-PK antibodies, but not with non-immune IgG used as a negative control (Figure 1, lanes 1, 2).
We next examined levels of c-RET protein expression in NHEM and human malignant melanoma cells (G361, HM3KO and MNT-1) (lanes 2 5 in Figure 5).
Anti-CD3 ligation increased the level of precipitated RasGRP-1 (lane 6), as did anti-LFA-1 (lanes 7) and a combination of anti-CD3/LFA-1 (lane 8).
Exogenous Jak2 1-525) was unable to phosphorylate the GHR upon GH addition, while Jak2(Y119E) induced a very low level of phosphorylation as compared to wild-type Jak2, probably due to a low residual affinity of the mutant Jak2 for box-1 (lanes 4, 6, 8), indicating that Jak2-GHR binding is essential for GHR phosphorylation (Fig. 1C).
