Full size image To evaluate the involvement of SCD1 in endothelial ciliation, an IP-ABE assay was performed to determine the effect of SCD1 on ARL13B palmitoylation. The results showed that SCD1 overexpression induced a substantial reduction in ARL13B palmitoylation in HUVECs, which was effective...
4i,j). Additionally, pulse-chase assays using radio-labelled fatty acids to trace degradation of TG showed that spartin KO cells had ~48% lower rates of TG degradation during 3 h after OA withdrawal (Fig. 4k). A similar impairment of TG degradation was found in cells lacking a core ...
4i,j). Additionally, pulse-chase assays using radio-labelled fatty acids to trace degradation of TG showed that spartin KO cells had ~48% lower rates of TG degradation during 3 h after OA withdrawal (Fig. 4k). A similar impairment of TG degradation was found in cells lacking a core ...
This relatively simple assay can be performed on fresh, frozen or frozen-lyophilized sediment samples with chloroform methanol extraction and subsequent phosphate determination. The sedimentary lipid phosphate recovery correlates with the extractible ATP and the rate of DNA synthesis. Pulse-chase ...
Pulse-chase experiments showed that apoC-III expression promoted VLDL1 secretion even under conditions where the MTP activity was inhibited immediately after the formation of lipid-poor apoB-100 particles, suggesting an involvement of apoC-III in the second-step VLDL assembly process. Consistent with...
Detailed protocols have been described (also see Experimental Procedures) for purification of EPs and LPs from Dictyostelium using a pulse-chase strategy (Barak et al., 2014, Gotthardt et al., 2006). Phagosomes purified from Dictyostelium were used for in vitro motility assays (Figure 1B; ...
Pulse-chase assay of BODIPY 558/568 C12 (Red C12) HEK293 cells were pulsed by an incubation in complete medium (DMEM with 10% fetal bovine serum) containing 1 μM BODIPY 558/568 C12 (ThermoFisher Scientific, D3835) for 16 h. Cells were washed with PBS three times and then chased...
BODIPY 558/564 Red C12 pulse-chase assay The BODIPY 558/564 Red C12 pulse-chase assay was performed as previously described29. In this experiment, HK-2 cells cultured in DMEM/F12 supplemented with 10% FBS containing 1 μM BODIPY 558/564 Red C12 were chased for 16 h in the absence...
B: “Pulse-chase” experimental design. Cells were incubated with “pulse” media containing [1-14C] oleate, washed, and stimulated with forskolin. Metabolism was assessed in the 4 h “chase” period. C: 14C fatty acid incorporated into triglyceride (TAG), presented as a percentage of total...
13 The cellular recycling of LNP-siRNA was determined by measuring the intracellular and extracellular amount of siRNA over time using the pulse-chase protocol outlined in the “Methods” section. As shown in Figure 3, in the absence of NP3.47, ∼60% of the previously internalized siRNA was...