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: International Journal of Pharmaceutics: X

Article Title: Roadmap for the accelerated development and clinical translation of fluorescent tracers: Adalimumab-680LT as a proof of concept

doi: 10.1016/j.ijpx.2026.100514

Figure Lengend Snippet: Results for the feasibility testing and lab development of adalimumab-680LT . SE-HPLC chromatograms of (A) unmodified adalimumab and (B) adalimumab-680LT at 280 nm, (C) chromatogram of adalimumab-680LT and free IRDye 680LT at 676 nm and (D) results of the indirect ELISA of unmodified adalimumab and adalimumab-680LT.

Article Snippet: Briefly, to remove excipients in the solution and to optimise the pH for labelling, adalimumab registered product (Humira®, AbbVie) was buffer exchanged to a 50 mM sodium phosphate buffer pH 8.5 (Apotheek A15, Gorinchem, the Netherlands) using pre-equilibrated PD-10 columns (Cytiva lifesciences, Chicago, IL, USA). cGMP grade IRDye 680LT NHS-ester (LI-COR Biosciences, Lincoln, NE, USA), dissolved in dimethyl sulfoxide (DMSO) (Sigma Aldrich, Darmstadt, Germany) 5 mg/mL, was added to the adalimumab solution in a molar dye-to-protein ratio of 2:1.

Techniques: Indirect ELISA

Journal: International Journal of Pharmaceutics: X

Article Title: Roadmap for the accelerated development and clinical translation of fluorescent tracers: Adalimumab-680LT as a proof of concept

doi: 10.1016/j.ijpx.2026.100514

Figure Lengend Snippet: Stability results of adalimumab-680LT. Release specifications are displayed with dotted lines, and end of shelf-life specifications are displayed with dashed lines. (A) Protein concentration, (B) percentage of free dye, (C) percentage of aggregates, and (D) target binding affinity of lab run 1 and 2 were tested during 3 months. For the technology transfer batch, (E) protein concentration, (F) percentage of free dye, (G) percentage of aggregates and (H) target binding affinity were tested at two different temperatures during 18 or 24 months. The stability study of the technology transfer batch at 2–8 °C is still ongoing. (A-C) are means of two different measurements, ( E -G) are means + standard deviations of three different measurements, and (D and H) are means of two measurements.

Article Snippet: Briefly, to remove excipients in the solution and to optimise the pH for labelling, adalimumab registered product (Humira®, AbbVie) was buffer exchanged to a 50 mM sodium phosphate buffer pH 8.5 (Apotheek A15, Gorinchem, the Netherlands) using pre-equilibrated PD-10 columns (Cytiva lifesciences, Chicago, IL, USA). cGMP grade IRDye 680LT NHS-ester (LI-COR Biosciences, Lincoln, NE, USA), dissolved in dimethyl sulfoxide (DMSO) (Sigma Aldrich, Darmstadt, Germany) 5 mg/mL, was added to the adalimumab solution in a molar dye-to-protein ratio of 2:1.

Techniques: Protein Concentration, Binding Assay

Journal: Cell Reports Medicine

Article Title: An inhalable microbe-oncolytic virus consortium for lung cancer treatment

doi: 10.1016/j.xcrm.2026.102771

Figure Lengend Snippet: In vivo distribution and efficacy assessment in orthotopic lung cancer mice model (A) In vivo imaging of the internal distribution of the consortium, PEG-Ads, and Ads at tumor sites, with comparison to intravenous administration ( n = 3). (B) Quantitative analysis of average fluorescence intensity per unit area corresponding to (A), showing lung deposition over time ( n = 3). (C) Distribution of formulations in excised organs, observed via IVIS at 24 h post-administration following euthanasia ( n = 3). (D) Quantitative chart of fluorescence intensity per unit area for each organ ( n = 3). (E) Quantitative fluorescence PCR analysis of Ad retention in lung tissues 12 h post-administration. (F) CLSM images of lung sections showing distribution of the consortium, PEG-Ads, and Ads, with lung tissues stained by DAPI and Ads labeled with FITC-NHS ester (one representative section from three mice is shown). (G) Tumor growth and body weight monitoring in TC-1-hCD46-Luc tumor-bearing mice treated with intratracheal nebulization of formulations. Terminal procedures, including organ collection for immune analysis, were conducted on day 10. (H) IVIS analysis of tumor bioluminescence and fluorescence intensity per unit area in different treatment groups, demonstrating treatment efficacy ( n = 6). Statistical analysis: data are presented as mean ± SD. Statistical significance was calculated using two-tailed Student’s t test (∗∗∗∗ p < 0.0001).

Article Snippet: FITC-NHS ester , Macklin (Shanghai, China) , Cat# C804900.

Techniques: In Vivo, In Vivo Imaging, Comparison, Fluorescence, Staining, Labeling, Two Tailed Test