Journal: The Journal of Cell Biology

Article Title: TIP47 functions in the biogenesis of lipid droplets

doi: 10.1083/jcb.200812042

Figure Lengend Snippet: TIP47 and MPR46 localize to different compartments. (A and B) Staining of HeLa cells for MPR46 (green), TIP47 (red), and neutral lipid (blue) at steady state (A) or after oleic acid feeding (B). Arrows point to small TIP47-positive structures. (inset) Western blot for the indicated proteins after fractionation of cells into membranes (M) and cytosol (C). Values are shown in kilodaltons. (C–K) Staining of MPR46, TIP47, and the indicated sorting proteins in HeLa cells at steady state (C–H) or after oleic acid feeding (I–K). Colocalization is indicated in yellow. Bars, 15 µm.

Article Snippet: DNA was transfected into HeLa cells using Effectene (QIAGEN) according to the manufacturer's instructions.

Techniques: Staining, Western Blot, Fractionation

Journal: The Journal of Cell Biology

Article Title: TIP47 functions in the biogenesis of lipid droplets

doi: 10.1083/jcb.200812042

Figure Lengend Snippet: Endogenous TIP47 and Rab9 do not colocalize. HeLa cells were fixed and stained for the indicated proteins. GFP-Rab7 and GFP-Lamp1 were transiently transfected 48 h prior to fixation. Colocalization is indicated in yellow. The inset displays a cell with small LDs at steady state. Bar, 15 µm.

Article Snippet: DNA was transfected into HeLa cells using Effectene (QIAGEN) according to the manufacturer's instructions.

Techniques: Staining, Transfection

Journal: The Journal of Cell Biology

Article Title: TIP47 functions in the biogenesis of lipid droplets

doi: 10.1083/jcb.200812042

Figure Lengend Snippet: CFP-Rab9 does not colocalize with YFP-TIP47. (A–H) HeLa cells were transiently transfected with YFP-TIP47 (A and E, green), CFP-Rab9 (B and F, green), CFP-Rab9, and YFP-TIP47 (C and G; green, CFP; red, YFP) or with CFP-ADRP and YFP-TIP47 (D and H; green, CFP; red, YFP). Cells depicted in the bottom panels were fixed after oleic acid (OA) feeding. A neutral lipid stain is shown in red (E and F) or blue (G and H). Brightness and contrast of TIP47 staining in A and E were enhanced to visualize the otherwise faint cytoplasmic staining. Colocalization is indicated in yellow. Bar, 15 µm.

Article Snippet: DNA was transfected into HeLa cells using Effectene (QIAGEN) according to the manufacturer's instructions.

Techniques: Transfection, Staining

Journal: The Journal of Cell Biology

Article Title: TIP47 functions in the biogenesis of lipid droplets

doi: 10.1083/jcb.200812042

Figure Lengend Snippet: TIP47 KD does not affect MPR localization and endocytosis. (A–F) Mock-transfected HeLa cells (control; A and B), TIP47 KD (C and D), and Vps35/26 KD cells (E and F) were either stained for MPR300 (red) at steady state (A, C, and E) or allowed to internalize anti-MPR300 antibodies (B, D, and F) before fixation and detection of uptake (red) and staining of TGN46 (green). Colocalization is indicated in yellow. The RNAi efficiency is shown in . Bar, 15 µm.

Article Snippet: DNA was transfected into HeLa cells using Effectene (QIAGEN) according to the manufacturer's instructions.

Techniques: Transfection, Control, Staining

Journal: The Journal of Cell Biology

Article Title: TIP47 functions in the biogenesis of lipid droplets

doi: 10.1083/jcb.200812042

Figure Lengend Snippet: The 11-mer repeat region of TIP47 is sufficient for LD targeting. Variants of human TIP47 fused to a 6×His tag or to GFP were expressed in HeLa cells followed by analysis of LD recruitment after oleic acid feeding. The localization was classified as +++ if all protein was visible on the LD surface (A). ++ indicates LD labeling as well as cytoplasmic detection (B). −/+ indicates a dominant localization in the cytoplasm. − indicates lack of any LD staining (C). wt, wild type.

Article Snippet: DNA was transfected into HeLa cells using Effectene (QIAGEN) according to the manufacturer's instructions.

Techniques: Labeling, Staining