Journal: bioRxiv

Article Title: Microenvironmental Determinants of Reaction Kinetics in Biomolecular Condensates Probed with Protein Ligation

doi: 10.64898/2026.03.26.714449

Figure Lengend Snippet: The presence of unreacted fractions and the heterogeneous clustering of client molecules under certain crowded conditions. ( A ) Reaction conversion over time in a bulk solution of 20 µM clients versus solutions with high concentrations of PEG. Bold data points in all conversion plots denote the average conversion at each time point, with raw data depicted as semi-transparent dots. The solid line indicates the best fit to modified second-order kinetics, and dashed lines represent the fitted unreacted fraction. ( B ) Confocal microscopy images of the reaction clients (mCh in red, GFP in green). In 50% PEG400, the clients associate and compartmentalize, and this phase separation is reversible upon dilution. Scale bars, 100 µm. ( C ) Reaction kinetics and unreacted fractions within three types of condensates (PRM–SH3, LAF, and FUS), where C client represents the internal client concentration within the condensate phase. ( D ) Condensate FRAP curves for PRM–SH3 and LAF scaffolds, alongside the recovery of mCh client partitioned within LAF condensates. Error bars represent ±1 standard deviation (SD; N=3). ( E ) Representative FRAP images for PRM–SH3, LAF, and FUS condensates. Scale bars, 10 µm. ( F ) Confocal microscopy images showing that the heterogeneous clustering of mCh clients becomes more pronounced as the unreacted fraction increases within the condensates. Scale bars, 10 µm.

Article Snippet: The samples were then centrifuged at 4000 rpm for 10 min (to sediment the condensates) and imaged by confocal laser microscopy (A1R HD25 with Eclipse Ti2, Nikon) using a 60×/1.40 numerical aperture (NA) apochromatic oil-immersion objective lens.

Techniques: Modification, Confocal Microscopy, Concentration Assay, Standard Deviation

Journal: Molecular and cellular endocrinology

Article Title: GDF15 inhibits early-stage adipocyte differentiation by enhancing HOP2 expression and suppressing C/EBPα expression.

doi: 10.1016/j.mce.2025.112461

Figure Lengend Snippet: Fig. 2. GDF15 expression is reduced by lysosomal degradation via the autophagic pathway. (A) Changes in GDF15 protein levels upon autophagic inhibition. ADSCs were treated with autophagy inhibitors, including chloroquine (CQ, 50 μM), ammonium chloride (NH4Cl, 25 mM), leupeptin (200 μM), pepstatin A (100 μM) + E−64 (35 μM), for 24 h prior to performing western blotting. (B, D) ADSCs were pre-treated with CQ (50 μM) for 1 h, and after changing the medium to a differentiation medium (DM), were cultured for 16 h and then fixed. Subsequently, the cells were treated with (B) antibodies against GDF15 (green) or SQSTM1 (red) and (D) antibodies against GDF15 (green) or LAMP1 (red) prior to capturing images using a confocal microscope. Yellow = merge/co-localization; scale bars: 10 μm. (C, E) Quantification of the co-localization between GDF15 and MAP1LC3B (C) and GDF15 and LAMP1 (E). Merged images in (B and D) were assessed for GDF15: MAP1LC3B (B) and GDF15:LAMP1 (D) co-localization using NIS Elements software and Pearson correlation analysis. Data are presented as the means ± standard deviation of 10 randomly selected cells for each treatment; ***P < 0.001. (F–H) Changes in GDF15 expression induced by knockdown of autophagy-related genes: ATG5 (F), ATG16L (G), and SQSTM1 (H). After transfection with scrambled siRNA or ATG5-, ATG16L-, and SQSTM1-specific siRNA in ADSCs, the medium was changed to DM and cultured 24 h later. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Images of the stained cells were acquired using a confocal laser microscope (A1 R HD25 NSIMS; Nikon, Tokyo, Japan) at the three-dimensional immune system imaging core facility of Ajou University.

Techniques: Expressing, Inhibition, Western Blot, Cell Culture, Microscopy, Software, Standard Deviation, Knockdown, Transfection