Journal: Kidney360

Article Title: Differential Assembly of Native Epithelial Sodium Channel Complexes Across Mouse Epithelial Tissues

doi: 10.34067/KID.0000001261

Figure Lengend Snippet: Immunofluorescence staining of kidney tissue from homozygous ENaC γ -VF mice. (A) Schematic of the tagged ENaC γ subunit produced from the ENaC γ -VF allele, which includes a C-terminal mVenus, 3C protease site, and 3xFLAG tag. (B) Representative immunofluorescence image showing colocalization of mVenus and ENaC γ in kidney tissue. mVenus signal was detected using a FITC-conjugated anti-GFP primary antibody (goat, Rockland, 1:100) and Alexa Fluor 488 donkey anti-goat secondary antibody. Endogenous ENaC γ was detected using a rabbit anti-ENaC γ primary antibody (StressMarq, 1:500) targeting the C-terminus, and Alexa Fluor 555 donkey anti-rabbit secondary antibody. (C) Close-up view highlighting localization of tagged ENaC γ . Anti-GFP, anti-green fluorescent protein; DAPI, 4′,6-diamidino-2-phenylindole; ENaC, epithelial sodium channel; ENaC γ -VF, epithelial sodium channel γ - m Venus FLAG.

Article Snippet: Primary antibodies included FITC-conjugated goat anti-green fluorescent protein (anti-GFP; Rockland, 1:100) for mVenus detection, rabbit anti-rat ENaC γ (Stressmarq, 1:500), mouse anti-Aquaporin-2 (Santa Cruz, 1:50), rabbit anti-ENaC α (Loffing Lab, 1:500), and chicken anti-phosphorylated sodium-chloride cotransporter (Welling Lab, 1:5000).

Techniques: Immunofluorescence, Staining, Tissue, Produced

Journal: bioRxiv

Article Title: Local fluidization of an active cytoplasmic gel partitions large cells

doi: 10.64898/2026.08.11.744254

Figure Lengend Snippet: a , Immunofluorescence images of a Xenopus laevis zygote fixed after the first mitosis, viewed from the animal pole. CPC (anti-INCENP-AF647) localized to the boundary between sister asters (microtubules imaged with anti-tubulin-AF488) and co-localized with an ER-depleted zone (ER labeled with anti-LNPK-AF568). b , Live images of a medaka embryo during 2-cell to 4-cell stage, showing dynamics of microtubules (HiLyte 647-tubulin), CPC (mCherry-Ol-Aurora B) and F-actin (Lifeact-EGFP) from mitosis to cytokinesis. c , CPC (anti-INCENP-AF647) localization in Xenopus egg extract co-imaged with microtubules (EB1-mApple), F-actin (Lifeact-GFP), and mitochondria (MitoView 405) at late anaphase. Top: untreated. Bottom: treated with 0.5 µM barasertib-HQPA. d , Example trajectories of two PEG-coated Fluoresbrite beads of 0.5 µm diameter in particle-tracking microrheology. Scale bar, 5 µm. e , Sampled areas in anaphase cycling egg extract with different cytoskeletal composition. f , Ensemble-averaged pairwise mean-squared displacement (MSD pair ) as a function of time lag τ for particles within each of the three indicated areas in e . g , Apparent diffusion coefficient (D) calculated from MSD pair between adjacent beads. For actin-intact controls, 8 areas (65x65 µm 2 ) in each category were sampled from independent experiments with 3 different egg extracts. The same extracts were used to prepare latrunculin B-added samples, where 20 areas were sampled in microtubule asters and cytoplasm, respectively. Data are mean ± SD. Also see . h , Local D calculated within overlapping 42x42 µm 2 grids, locally averaged, and overlaid onto microtubule images (EB1-mApple). Examples shown are microtubule asters growing against microtubule-free cytoplasm, in actin-intact egg extract (top) or extract treated with 10 µM LatB (bottom). Also see . All scale bars except d , 200 µm.

Article Snippet: HiLyte 647-tubulin (Cytoskeleton, Inc. TL670M) and mRNAs were mixed with final concentrations of 800 ng/μL for tubulin and150 ng/μL for each mRNA.

Techniques: Immunofluorescence, Labeling, Diffusion-based Assay