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. 2012 Sep 17;21(24):5318–5328. doi: 10.1093/hmg/dds388

Figure 4.

Figure 4.

Transient transfection of ATGL mutant proteins in HeLa cells. (A) HeLa cells, loaded for 18 h with OA (400 µm) complexed to BSA (6:1 molar ratio), were transiently transfected with one of the following plasmids: pEGFP, pATGL-EGFP, pATGL(Ser47Ala)-EGFP, pATGL(Asp166Gly)-pEGFP, pATGL(Asn172Lys)-EGFP, pATGL(Pro195Leu)-EGFP, pATGL(Arg221Pro)-EGFP and pATGL(Gly483Arg)-EGFP. After 24 h, the cells were fixed and stained with ORO. Fluorescence of EGFP and ATGL-EGFP fusion proteins is in green. Scale bar: 10 µm. (B and C) ORO-stained LDs of Hela cells non-transfected and transfected with pEGFP (V), pATGL-EGFP (WT), pATGL(Ser47Ala)-EGFP (M0), pATGL(Asp166Gly)-pEGFP (M1), pATGL(Asn172Lys)-EGFP (M2), pATGL(Pro195Leu)-EGFP (M3), pATGL(Arg221Pro)-EGFP (M4) and pATGL(Gly483Arg)-EGFP (M5) plasmids. The cells were analyzed for number (b) and dimension (c) (cross-section area) of LDs per cell. The values represent the means of 80–95 cells for each transfected mutant. Experiments were run in triplicate.