Journal: Science Advances

Article Title: Reconstituted nascent adhesion condensates drive actin polymerization on supported lipid bilayers

doi: 10.1126/sciadv.aeb6691

Figure Lengend Snippet: ( A ) Pyrene-actin polymerization assay in the absence or presence of profilin, CapZ, and/or zyxin-VASP or vinculin-VASP condensates. Error bars represent the standard deviations of three independent experiments. ( B ) Schematic of the experimental setup for condensate-mediated actin polymerization on SLBs. ( C and D ) Representative images showing G-actin partitioning into zyxin-VASP (C) and vinculin-VASP (D) condensates. Scale bars, 10 μm. ( E ) Time-lapse image series showing actin polymerization by zyxin-VASP and vinculin-VASP condensates. Scale bars, 1 μm. ( F ) Representative images of actin filaments nucleating outside condensates; arrows indicate filament origin sites, and circles mark condensate positions. Scale bars, 1 μm. ( G ) Box plot quantifying the fraction of actin filaments originating from condensates compared to those originating outside condensates. Boxes indicate the 25th to 75th percentiles, the median is shown as a line, and whiskers represent 1.5× the interquartile range; P < 0.05 is reported. Significance was tested with a Mann-Whitney U test. Each condition contains n measurements from three independent experiments. Zyxin is visualized with zyxin-mCherry, and actin is visualized with 12.5% actin-ATTO488. Condensates are imaged by epifluorescence microscopy and actin filaments by TIRF microscopy.

Article Snippet: ATTO488 NHS-ester (FP201-488) was purchased from Jena Bioscience.

Techniques: Polymerization Assay, MANN-WHITNEY, Epifluorescence Microscopy, Microscopy

Journal: Science Advances

Article Title: Reconstituted nascent adhesion condensates drive actin polymerization on supported lipid bilayers

doi: 10.1126/sciadv.aeb6691

Figure Lengend Snippet: ( A ) Representative image of an actin network generated by zyxin-VASP condensates after 20 min, showing prominent actin bundles. Scale bar, 10 μm. ( B ) Representative image of an actin network generated by vinculin-VASP condensates after 20 min, showing predominantly single filaments. Scale bar, 10 μm. ( C ) Time-lapse image series showing zyxin-VASP–generated filaments forming a bundle upon filament-filament encounter; arrows mark the interaction point. Scale bar, 1 μm. ( D ) Time-lapse image series showing vinculin-VASP–generated filaments crossing over each other upon encounter; arrows mark the interaction point. Scale bar, 1 μm. ( E ) Box plot quantifying actin bundle line density (number of bundles intersecting a line). Boxes indicate the 25th to 75th percentiles, the median is shown as a line, and whiskers represent 1.5× the interquartile range; P < 0.05 is reported. Significance was tested with a Mann-Whitney U test. Each condition contains n measurements from three independent experiments. Actin is visualized with actin-ATTO488. Actin images are imaged using TIRF microscopy.

Article Snippet: ATTO488 NHS-ester (FP201-488) was purchased from Jena Bioscience.

Techniques: Generated, MANN-WHITNEY, Microscopy

Journal: Science Advances

Article Title: Reconstituted nascent adhesion condensates drive actin polymerization on supported lipid bilayers

doi: 10.1126/sciadv.aeb6691

Figure Lengend Snippet: ( A and B ) Time-lapse image series illustrating changes in condensate shape upon actin polymerization. Scale bars, 5 μm. ( C and D ) Histograms showing changes in condensate eccentricity as a function of actin polymerization. ( E ) Representative FRAP image series of zyxin-VASP and vinculin-VASP condensates. Scale bars, 1 μm. ( F ) Normalized FRAP recovery curves and extracted parameters [fast half-time of recovery ( t 1 / 2 _ fast ) and mobile fraction] for zyxin-VASP and vinculin-VASP condensates. Data are shown as the means ± standard deviation. Error bars represent the standard deviations of three independent experiments. ( G ) Representative image of an actin network generated by zyxin-VASP and subsequently contracted by NMMII in the presence of α-actinin after 60 min, showing clustered actin filaments and condensates. Scale bar, 10 μm. ( H ) Time-lapse image series showing actin network contraction and concomitant clustering of zyxin-VASP condensates in the presence of NMMII and α-actinin. Scale bar, 5 μm. ( I ) Box plot quantifying the number of zyxin-VASP condensates before and after actin network contraction in the presence of NMMII and α-actinin. Boxes indicate the 25th to 75th percentiles, the median is shown as a line, and whiskers represent 1.5× the interquartile range. Significance was tested with a Mann-Whitney U test. Each condition contains n measurements from three independent experiments. Zyxin is visualized with zyxin-mCherry, vinculin is visualized with 10% vinculin-AF555, and actin is visualized with actin-ATTO488. Condensates are imaged using epifluorescence microscopy, and actin filaments are imaged using TIRF microscopy.

Article Snippet: ATTO488 NHS-ester (FP201-488) was purchased from Jena Bioscience.

Techniques: Standard Deviation, Generated, MANN-WHITNEY, Epifluorescence Microscopy, Microscopy

Journal: bioRxiv

Article Title: Dual Pathways of Extracellular ATP Action in Cancer Cells: Purinergic Signaling–Driven Senescence and Macropinocytic ATP Internalization

doi: 10.64898/2026.04.23.720363

Figure Lengend Snippet: (A–B) Short-time interval ATP concentration assay (minutes). Intracellular ATP (iATP, % of control) for A549 (A) and A549STC1KO (B) cells measured at the indicated induction times between 30-120 minutes. (C–D) Long-time interval ATP concentration assay (hours). iATP (%) for A549 WT (C) and STC1-KO (D) across induction times between 2-4 hours. (E) Immunofluorescence/confocal imaging of macropinocytosis. A549 WT, A549 STC1-KO (two independent clones), and NL20 control cells were stained with NHF-ATP (green; ATP analog), High molecular weight dextran (red-macropinosome marker), and DAPI (blue; cell nuclei). Merged images show colocalization (yellow). STC1-KO cells display visibly reduced FITC-dextran uptake relative to WT based on gene expression reduction, while NL20 shows minimal uptake. Scale bars: 20 µm (A549 and STC1-KO), 10 µm (NL20). Data is representation for one set of biological replicates as mean ± SD for (n ≥ 5 technical replicates). Colors denote the eATP dose as indicated in the in-plot legend. One-way ANOVA with post-hoc multiple-comparisons; *P < 0.05,**P < 0.01,***P < 0.001, ****P < 0.0001.

Article Snippet: For fluorescence microscopy, NHF-ATP was obtained from Jena Bioscience (NU-810-488), ProLong TM Gold Antifade Mountant with DAPI from Thermo Fisher and high molecular weight fluorescent TMR-dextran (HMWFD) 70,000 Daltons MW, Neutral (no electric charge) from Invitrogen (D818).

Techniques: Concentration Assay, Control, Immunofluorescence, Imaging, Clone Assay, Staining, High Molecular Weight, Marker, Gene Expression

Journal: bioRxiv

Article Title: Dual Pathways of Extracellular ATP Action in Cancer Cells: Purinergic Signaling–Driven Senescence and Macropinocytic ATP Internalization

doi: 10.64898/2026.04.23.720363

Figure Lengend Snippet: Tumor xenografts were established by subcutaneous injection of 1×10 –5×10 cells into the right flank of 3–4-week-old male NU/J nude mice. When tumors reached ∼200–500 mm³, tumors were injected intratumorally with DMEM control, HMWFD tracer, or HMWFD plus NHF-ATP (100 μM) in a total volume of 50 μL using 27G needles. Tumors were harvested within ∼7–8 minutes post-injection, embedded in OCT, and cryosectioned (10 μm). Sections were ethanol-fixed, PBS-washed, and mounted with antifade medium containing DAPI. Images were acquired using a Nikon NiU epifluorescence microscope with identical exposure settings across groups and processed using Nikon NIS-Elements software. A-H. NHF-ATP and HMWFD colocalize in HT29 (A), PANC1 (B), MCF7 (C), SK-HEP1 (D), HOP-92 (E), H1299 (F), and A549 (G) and A375 (H). respectively. Color: Blue:DAPI (nuclei), Red: HMWFD, Green: NHF-ATP, Yellow: Co-localization. Scale bar: 10 µm, 100x magnification.

Article Snippet: For fluorescence microscopy, NHF-ATP was obtained from Jena Bioscience (NU-810-488), ProLong TM Gold Antifade Mountant with DAPI from Thermo Fisher and high molecular weight fluorescent TMR-dextran (HMWFD) 70,000 Daltons MW, Neutral (no electric charge) from Invitrogen (D818).

Techniques: Injection, Control, Microscopy, Software