|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
|
Fig. 5. Effect of AGAP2 on endosomal markers. (A) Colocalization of AGAP2 with endosomal markers. (A, a-d) AGAP2 colocalized with AP-1 and transferrin receptor. FLAG-AGAP2 under the control of SV40 promoter (in pSI vector) was transfected into HeLa cells for 24 hours. The cells were fixed and stained for AP-1 and transferrin receptors (TfnR). The overexpressed AGAP2 was detected by staining for the FLAG tag. Colocalization of AGAP2 with AP-1 and TfnR was indicated by arrows. (A, e-j) Colocalization of AGAP2 with Rab4. FLAG-AGAP2 in pSI vector was transfected with GFP-Rab4, GFP-Rab5 and GFP-Rab11 into HeLa cells for 24 hours. The overexpressed AGAP2 was detected by staining for the FLAG tag. AGAP2 colocalized with Rab4 (arrows in e, f), but not with Rab5 (g,h) or Rab11 (i,j). (B) Effect of AGAP2 on endogenous Rab4 and TfnR. HeLa cells were transfected with FLAG-AGAP2 driven by the CMV promoter (in pCI vector) for 24 hours. Endogenous Rab4 (B, a,b), Rab11 (B, c,d) and transferrin receptors (B, e,f) were visualized by staining with specific antibodies. Transfected cells were visualized by staining for the FLAG tag and indicated by arrows. (C) Relative expression level of [FLAG]AGAP2. HeLa cells were transfected with empty vector, pSI-[FLAG]AGAP2 or pCI-[FLAG]AGAP2 for 24 hours. Cells were harvested and lysed. The lysates were subjected to SDS-PAGE and western blot using polyclonal anti-FLAG antibody.
|
