Journal: bioRxiv

Article Title: PKD2 structural destabilization drives primary cilia degeneration and ADPKD pathogenicity

doi: 10.64898/2026.06.24.734313

Figure Lengend Snippet: A ) Left , image of a voltage clamped primary cilia in the on-cilia configuration. The primary cilium of HEK (PKD1 Null :PKD2 Null ) cells transiently expressing PKD2 channels are illuminated by ARL13B-GFP under 475 nm laser excitation. Right , example single channel recordings from the cilia membrane activated by depolarizing voltage steps. Note, no open channel events were observed in cilia patch recordings from cells expressing D511V channels. B ) Left , example polycystin unitary single channel events recorded from cilia membranes. Right , corresponding histograms indicating the frequency of a variable simultaneous open channel events (O 1 -O 3 ) and closure (C). Note, no channel openings are detected from cilia recordings from cells expressing D511V channels. C ) Column plots indicating the frequency of the maximum simultaneous open channel events from cilia patches. Number of events are indicated in parenthesis. D ) Image of a voltage clamped giant unilaminear vesicals (GUVs) reconstituted with cell free expressed PKD2-GFP protein. E ) Top , example WT PKD2-GFP unitary single channel events recorded (HP = 40mV) from GUVs composed of either 1,2-diphytanoyl-sn-glycero-phosphocholine (DPhPC) or 95% DPhPC and 5% cholesterol (Bovine). Bottom , lack of single channel events from D511V channels reconstituted in GUVs. F ) Column plots indicating the frequency of the maximum simultaneous open channel events from GUVs patches. Number of recordings indicated above for each column.

Article Snippet: Lipids DPhPC (95%) and cholesterol (bovine, 5%) were obtained from Avanti Polar Lipids (Alabaster, USA) and mixed in chloroform, where vacuum evaporation on glass formed small unilaminar vesicles (SUVs), rehydrated and passed through a 100-nm polycarbonate filter with a Mini-Extruder (Avanti Polar Lipids).

Techniques: Expressing, Membrane

Journal: bioRxiv

Article Title: Membrane curvature enhances lipid peroxidation in a composition-dependent manner

doi: 10.64898/2025.12.03.692219

Figure Lengend Snippet: A) Structure of DLPC and DPhPC. B) The fluorescence emission spectra of Laurdan for each lipid composition. C) The GP value of Laurdan for each lipid composition. D) The average fraction of oxidized C11-BODIPY over time for each diameter bin. E) The initial rate of oxidation as a function of vesicle diameter. The change in the fraction of oxidized C11-BODIPY after F) 10 minutes and G) 30 minutes of oxidation, relative to its initial value for each diameter bin. H) Representative fluorescence micrographs of MBLs before and after photobleaching (scale bar: 5 μm). I) Fluorescence recovery in MBLs over time. The red lines represent the lines of best fit from equation S7. J) Diffusion coefficients for each lipid composition. All vesicles used in panel B and C were composed of DLPC or DPhPC (99.8 mol%) and Laurdan (0.2 mol%). All vesicles used in panel D-G were composed of DPhPC (98.5 mol%), C11-BODIPY (0.5 mol%), DPPE-ATTO 647N (0.5 mol%), and DSPE-PEG(2000)-Biotin (0.5 mol%), and oxidation was induced using 7.5 μM H 2 O 2 and 0.15 μM FeSO 4 . The number of DPhPC SUVs in each diameter bin varied from 140–4959. Data points or bars in D-G represent the average value of the bin and the error bars correspond to the standard error of the mean for each bin. Each value displayed for a bin represents the midpoint of the bin. All bins were symmetric, with endpoints aligned to the start of the next bin, ensuring no overlap. MBLs used in panel H-J were composed of DLPC or DPhPC (99.875 mol%) and DPPE-ATTO 647N (0.125 mol%). Data points in I represent the average normalized fluorescence intensity. Bars in J represent the mean and the error bars correspond to the standard deviation ( N =5, unpaired, two-sample t-test, ****P<0.0001).

Article Snippet: DLPC (1,2-dilauroyl-sn-glycero-3-phosphocholine, 850335), DPhPC (1,2-diphytanoyl-sn-glycero-3-phosphocholine, 850356), POPC (1-palmitoyl-2-oleoyl-glycero-3-phosphocholine, 850457), DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine, 850375), cholesterol (700100), DLiPC (1,2-dilinoleoyl-sn-glycero-3-phosphocholine, 850385), DSPE-PEG(2000)-Biotin (1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[biotinyl(polyethylene glycol)-2000] (ammonium salt), 880129) were purchased from Avanti Polar Lipids.

Techniques: Fluorescence, Diffusion-based Assay, Standard Deviation

Journal: bioRxiv

Article Title: Membrane curvature enhances lipid peroxidation in a composition-dependent manner

doi: 10.64898/2025.12.03.692219

Figure Lengend Snippet: A) Structure of DLiPC. B) Fluorescence recovery in SBLs over time. The lines represent the best fits with equation S7. C) Lateral diffusion coefficients for each membrane composition. D) The initial rate of oxidation as a function of vesicle diameter. The change in the fraction of oxidized C11-BODIPY after E) 5 minutes and F) 10 minutes of oxidation, relative to its initial value for each diameter bin. SBLs used in panel B and C were primarily composed of DOPC, 1:1 DLiPC:DPhPC (50% DLiPC), or 3:1 DLiPC:DPhPC (75% DLiPC), and contained 0.125 mol% DPPE-ATTO 647N. Data points in B represent the average normalized fluorescence intensity. Bars in C represent the mean and the error bars correspond to the standard deviation ( N =4 for DOPC, and N =5 for 50% DLiPC and 75% DLiPC, unpaired, two-sample t-test, ns: non-significance, *P<0.05). All vesicles used in panel D-F were primarily composed of DOPC, 1:1 DLiPC:DPhPC (50% DLiPC), or 3:1 DLiPC:DPhPC (75% DLiPC), and contained C11-BODIPY (0.5 mol%), DPPE-ATTO 647N (0.5 mol%), and DSPE-PEG(2000)-Biotin (0.5 mol%). Oxidation was induced using 7.5 μM H 2 O 2 and 0.15 μM FeSO 4 . The number of SUVs in each diameter bin varied from 320–4502 (50% DLiPC), and 263–3100 (75% DLiPC). Bars in D represent the average initial rate of oxidation calculated from linear regression of three time points (0, 5, and 10 minutes, solid fill) or slope of the first two time points (0 and 5 minutes, patterned fill) and the error bars correspond to the standard error calculated from linear regression (solid fill) or propagated error from the standard error (patterned fill). Bars in E and F represent the average value of the bin, and the error bars correspond to the standard error of difference. Each value displayed for a bin represents the midpoint of the bin. All bins were symmetric, with endpoints aligned to the start of the next bin, ensuring no overlap.

Article Snippet: DLPC (1,2-dilauroyl-sn-glycero-3-phosphocholine, 850335), DPhPC (1,2-diphytanoyl-sn-glycero-3-phosphocholine, 850356), POPC (1-palmitoyl-2-oleoyl-glycero-3-phosphocholine, 850457), DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine, 850375), cholesterol (700100), DLiPC (1,2-dilinoleoyl-sn-glycero-3-phosphocholine, 850385), DSPE-PEG(2000)-Biotin (1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[biotinyl(polyethylene glycol)-2000] (ammonium salt), 880129) were purchased from Avanti Polar Lipids.

Techniques: Fluorescence, Diffusion-based Assay, Membrane, Standard Deviation