pi(5)p dic4  (Echelon Biosciences)

Journal: bioRxiv

Article Title: The phosphoinositide PI5P impairs mitochondrial function through endosome-mitochondria proximity

doi: 10.1101/2025.09.22.677701

Figure Lengend Snippet: (A) Overexpression of IpgD induces mitochondrial fragmentation. Cells expressing myc-IpgD WT, C438S mutant or GFP-Inp54p were fixed and stained for mitochondria (left panel), and the mitochondrial fragmentation index was quantified across 3 independent experiments (right panel). Bar is 10µM in all figures unless otherwise stated. (B-C) Exogenous PI5P induces mitochondrial fragmentation and increases mitochondrial ROS (mtROS) production and decreased mitochondrial membrane potential (mtΔΨ). Cells were treated overnight with 15µM of PI5P or PI4P, fixed and stained for mitochondria (B) or incubated with MitoSOX (for mtROS production) or with TMRE (for mtΔΨ) and live-imaged (C). Quantifications from 3 independent experiments are shown on the right. (D) Bioenergetic profile of cells treated with PI5P. Treated cells were processed for a Mito Stress Test on a Seahorse flux analyser. A representative Seahorse trace including five technical replicates is shown. (E) Quantification of Basal respiration, Maximal respiration and Spare respiratory capacity from 3 independent experiments are shown. (F) Energy map from control or PI5P-treated cells showing a decrease in the oxidative capacity of the PI5P-treated cells, both in basal and stressed (FCCP treated) conditions. (G) Lysates from control or PI5P-treated cells were probed for the indicated antibodies (left panel). Quantifications are shown on the right. Statistical significance was determined by one-way ANOVA with Tukey’s post hoc analysis for multiple comparisons or unpaired t-test. *p<0.05, **p < 0.01, ***p < 0.001,****p < 0.0001, ns non-significant.

Article Snippet: Cells were treated with STA (100nM), DSMO (vehicle only) or cell permeant di-C4 PI5P (P-5004, Echelon Biosciences) or PI4P (P-4004, Echelon Biosciences) as indicated.

Techniques: Over Expression, Expressing, Mutagenesis, Staining, Membrane, Incubation, Control

Journal: bioRxiv

Article Title: The phosphoinositide PI5P impairs mitochondrial function through endosome-mitochondria proximity

doi: 10.1101/2025.09.22.677701

Figure Lengend Snippet: (A) H9C2 cells were fixed and stained for mitochondria (in green) or for PI5P (in green). Zoomed region (i) and (ii) are shown below. (B) Line scan of fluorescence intensity along each arrow from inserts (i) and (ii) from (A). (C) Proportion of mitochondrial vs. non-mitochondrial PI5P. Results are from 19 cells across 3 independent experiments. (D) Cells were treated with STA (100nM for the indicated time) or with DMSO (vehicle only), fixed and stained for mitochondria (in green) or PI5P (in red). Right panel show the quantification of PI5P fluorescence intensity in the mitochondrial (mito.) or non-mitochondrial mask (non-mito.). Results are from 13-27 cells across 3 independent experiments. Statistical significance was determined by unpaired t-test. *p<0.05, ***p < 0.001.

Article Snippet: Cells were treated with STA (100nM), DSMO (vehicle only) or cell permeant di-C4 PI5P (P-5004, Echelon Biosciences) or PI4P (P-4004, Echelon Biosciences) as indicated.

Techniques: Staining, Fluorescence

Journal: bioRxiv

Article Title: The phosphoinositide PI5P impairs mitochondrial function through endosome-mitochondria proximity

doi: 10.1101/2025.09.22.677701

Figure Lengend Snippet: (A) Cells were treated with 2DG (50mM) in complete medium for 4h, fixed and stained for mitochondria (in green) or PI5P (in red). Quantifications of total PI5P fluorescence, PI5P fluorescence intensity in the mitochondrial (mito.) or non-mitochondrial mask (non-mito.) from 3 independent experiments are shown on the bottom panel. (B) Cells were treated as in (A) except for a pre-treatment with STA (30min, 100nM) as indicated. Results are from 3 independent experiments. Statistical significance was determined by one-way ANOVA with Tukey’s post hoc analysis for multiple comparisons or unpaired t-test. *p<0.05, **p < 0.01, ***p < 0.001,****p < 0.0001.

Article Snippet: Cells were treated with STA (100nM), DSMO (vehicle only) or cell permeant di-C4 PI5P (P-5004, Echelon Biosciences) or PI4P (P-4004, Echelon Biosciences) as indicated.

Techniques: Staining, Fluorescence