Journal: iScience

Article Title: DACIT device for axon cancer cell interaction testing in 2D and 3D

doi: 10.1016/j.isci.2025.114557

Figure Lengend Snippet: 3D Spheroid invasion assay in DACIT (A) Experimental timeline of 3D spheroid invasion assays in DACIT. Tumor spheroids were added after axons reached the axonal compartment. (B) Representative images of 4T1 spheroids embedded in a collagen: Matrigel mix. Spheroid invasion was monitored in the presence of DMSO (control) or GM6001 (25 μM) at 2 h post-embedding (Day 0), 24 h (Day 1), and 48 h (Day 2). Scale bars, 200 μm. Graphs show the mean ± SEM relative spheroid area (C) and minimum-to-maximum circularity (D) of nine spheroids per treatment from two biologically independent experiments. Statistical differences (∗ p < 0.05 compared with control) were determined using an unpaired, two-tailed Mann-Whitney test. (E) Top (left) and side (right) views of tumor spheroids (4T1-mScarlet, magenta) embedded in a 3D ECM mix of collagen: Matrigel (blue) in the presence of neurons (yellow), fixed 24 h after invasion. Scale bars, 200 μm. (F) Brightfield image of a spheroid embedded in 3D ECM, 24 h post-embedding. Microgrooves are visible on the left side of the image. Scale bars, 200 μm. (G) Maximum projection of invasive strands from a 4T1-mScarlet 3D spheroid (magenta), 24 h post-embedding, interacting with axons (yellow, PGP9.5). Scale bars, 100 μm. (H) 3D Imaris reconstruction and (I) single z-slice of an invasive strand in the spheroid (magenta) interacting with axons (yellow). Scale bars, 50 μm.

Article Snippet: Scale bars, 100 μm. (H) 3D Imaris reconstruction and (I) single z-slice of an invasive strand in the spheroid (magenta) interacting with axons (yellow).

Techniques: Invasion Assay, Control, Two Tailed Test, MANN-WHITNEY

Journal: bioRxiv

Article Title: Alternative exon splicing reveals hidden mitochondrial targeting of PLIN3

doi: 10.64898/2026.02.18.706587

Figure Lengend Snippet: A) Representative electron micrographs of cells transfected with PLIN3B-P2A-BFP. Scale bar=500 nm. B) Representative electron micrographs of cells transfected with PLIN3A-P2A-BFP. Scale bar=500 nm. C) Representative immunocytochemistry structured illumination microscopy (SIM 2 ) images of mock transfected HeLa cells or HeLa cells transfected with PLIN3B-P2A-BFP. Cells were stained with MitoTracker Red CMXRos, anti-PLIN3B 4D72 antibody and with anti-TOMM20 antibody. Images were acquired with Elyra7 microscope. Scale bar=2.5 µm. N=3 independent experiments. D) Profile plot of dashed lines in . E) Correlation between electron micrograph and CLSM images of HeLa cells transfected with PLIN3B-P2A-BFB. The images correspond to the ROI in . White asterisk correspond to the same structure as in . Dashed ROIs corresponding to serial sections in . Scale bar=2 µm. F) Imaris 3D reconstruction of the ROI in in top and side view. Black dashed lines indicate the top and bottom serial sections in . Black arrow indicates serial sections direction. White and black asterisk correspond to the same structure as in and . G) Electron micrographs of serial sections from mitochondria in ROIs from . White asterisk corresponds to the same structure as in and . Scale bar=1 µm. H) CLSM image corresponding to section 4 in . White asterisk corresponds to the same structure as in . Cells were stained with MitoTracker Red CMXRos, PLIN3B 4D72 antibody and with anti-TOMM20 antibody.

Article Snippet: 3D reconstruction was performed with Imaris (Oxford Instruments, High Wycombe, UK, RRID:SCR_007370).

Techniques: Transfection, Immunocytochemistry, Microscopy, Staining

Journal: bioRxiv

Article Title: Two anchoring proteins control daughter apical complex assembly in Toxoplasma gondii

doi: 10.64898/2026.02.13.705759

Figure Lengend Snippet: a, Imaris 3D reconstruction of U-ExM images from syringed-lysed tachyzoites, either left untreated or IAA-treated for 24h. Tachyzoites were stained with anti-α/β tubulin (magenta) antibody and Hoechst (blue) to label microtubules and nuclear DNA, respectively. Images represent maximum-intensity projections of z-stack confocal sections with both side and top views (left and right panels, respectively). White arrows indicate mother apical complex. SPMT numbers annotated on the top-view tachyzoite images. Scale bars are 1μM. b, Representative images of apical markers in mother cells by U-ExM. U-ExM images of RCC1-2-mAID-HA 3 + CPH1-Ty2 (f) , + KinA-Ty2 (g) and + APR7-Ty2 (h) parasites, either left untreated or treated with IAA for 24h. Intracellular tachyzoites were stained with anti-Ty (green) and anti-α/β tubulin (magenta) antibodies to label Ty-tagged proteins and microtubules, respectively. Images represent maximum-intensity projections of z-stack confocal sections. White arrows indicate mother apical complex. Scale bars are 2μM. RCC1-2 depletion leads to loss of APR7 integrity. c, Gliding motility is impaired upon RCC1-2 depletion: Parasites were treated with A23187 to stimulate gliding motility visualized by gliding trails labeled with an SAG1 antibody. Scale bars are 5μM. d, RCC1-2 depletion is responsible of an invasion defect: Invasion was monitored for control (Tir1) and RCC1-2-depleted tachyzoites upon 24 h treatment with IAA. Values are reported as mean ± SD ( n = 3 biological replicates, each with three technical replicates). Statistical significance was determined by unpaired two-tailed Student’s t-test. e, Stimulated parasite egress is not affected in the absence of RCC1-2: Infected cells were treated with A23187 to stimulate parasite egress, measured as a number of egressed parasitophorous vacuoles over the total number. Egress was tested for control (Tir1) and RCC1-2-mAID-HA 3 cell lines in the absence of IAA, and upon 24h of IAA treatment. Values are reported as mean ± SD ( n = 3 biological replicates, each with three technical replicates). Statistical significance was determined by unpaired two-tailed Student’s t-test. At least 100 PVs were quantified in each replicate. P-values are non-significant for all datasets (two-tailed t-test). f, Representative images of egress assay treated as in e . Parasites were stained with anti-GAP45 (red) and anti-GRA3 (green) antibodies to label parasites plasma membrane and parasitophorous vacuoles, respectively. DNA is labeled with Hoechst (blue). Scale bars are 2μM.

Article Snippet: Supplementary movie 1: Imaris 3D reconstruction of U-ExM images from intracellular RCC1-2-mAID-HA 3 parasites.

Techniques: Staining, Labeling, Control, Two Tailed Test, Infection, Clinical Proteomics, Membrane