Fig 1: (A) Schematic of the microfluidic system to form a free-standing bilayer containing Proteo-LDs. (B) The secondary structure of ADRP was provided by the AlphaFold AI database. ADRP is reconstituted in the PC:PE monolayer covering the LD. (C) A bilayer is formed at the aperture between the channel and the cone, and LDs containing ADRP protein are introduced from the bottom channel and insert into the bilayer. (D) Schematic of the microfluidic device to form the free-standing monolayer. A phospholipid monolayer is formed at the buffer–triolein interface.
Fig 2: (A) Top view of a free-standing bilayer containing LDs. A side view of a 3D scan is provided in the top panel. The right panel provides the corresponding side view with only the protein channel. All the proteins are labeled in green (Alexa 488) and 2% (molar ratio) DOPE is labeled in red (Cy5). (B) 3D reconstructions of individual bilayer-embedded LDs for 0%, 20%, and 40% cholesterol, respectively (with 1 μM ADRP). For comparison, the 3D reconstructions of a bilayer-embedded LD with 0% cholesterol are given, and no ADRP was added. In the latter case, the neutral lipids were stained with BODIPY. (C) Scheme of the surface forces acting on a bilayer-embedded LD with an ideal lens shape (upper panel). Scheme of the surface forces acting on the bilayer (lower paner). (D) Comparison between wetting theory and the balance of surface forces as extracted from the symmetric 3D LD geometry.
Fig 3: (A) Series of fluorescence images showing a lipid monolayer (DOPC:DOPE, 3:1) between buffer and triolein before bleaching and during recovery. (Upper panel) DOPE-Cy5, 2% molar ratio; (Lower panel) ADRP-Alexa488, 1 μM. (B) FRAP curves extracted from fluorescence images, such as the one shown in (A) with the corresponding Soumpasis fits for DOPE-Cy5 (red dots) and ADRP-Alexa488 (green dots). Yellow dots correspond to the recovery of the DOPE-Cy5 fluorescence obtained by bleaching the entire bilayer-embedded LD surface, with same bilayer composition as in (A). (C) Diffusion coefficients of DOPE in the lipid monolayer, as a function of total lipid concentration and in the presence and absence of ADRP. The error bars are presented by the continuous lines. In (A–C), each value was obtained by averaging ≈ 30 different measurements.
Supplier Page from Abcam for Recombinant Human Perilipin-2 protein (denatured) (T7 tag N-Terminus)