Journal: Frontiers in Physiology

Article Title: A comprehensive approach to elucidating the pathophysiology of kidney fibrosis based on extracellular vesicle proteomics

doi: 10.3389/fphys.2026.1786999

Figure Lengend Snippet: TG2 suppression modulated proteins associated with ECM maintenance, inflammation, apoptosis, and TGF-β signaling in rTGF-β-treated hPTECs and NIH3T3 cells. (A) Western blots of fibronectin, periostin, collagen type IV, TNF-α, integrin-α5, and β-actin of hPTECs treated with rTGF-β and cysteamine (48 h), and (B) densitometric quantification normalized to β-actin. (C) Western blots of fibronectin, periostin, collagen type I-α1, integrin-α5, and β-actin of NIH3T3 cells treated with rTGF-β and cysteamine (24 h), and (D) densitometric quantification normalized to β-actin. (E) Light microscopic images of hPTECs and (F) NIH3T3 cells treated with rTGF-β and cysteamine. (G) Annexin V and propidium iodide (PI) staining of human proximal tubular epithelial cells (hPTECs) treated with rTGF-β and cysteamine (48 h), and (H) quantification. Statistical significance: * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001.

Article Snippet: The cells were incubated at 37 °C and 5% CO 2 for 48 h. NIH3T3 cells (ATCC: CRL-1658) were maintained in DMEM/F12 (Biowest, Riverside, MO, USA; Cat# L0092) supplemented with 10% FBS (Gibco, Cat# A4766801) and 1% 100X penicillin-streptomycin (Gibco, Cat# 15140-122).

Techniques: Western Blot, Staining

Journal: Nucleic Acids Research

Article Title: Repeat-rich RNA guides repetitive genomic elements into biomolecular condensates for heterochromatin organization and muscle integrity

doi: 10.1093/nar/gkag168

Figure Lengend Snippet: CU-rich RNA promotes heterochromatin condensate organization during differentiation. ( A ) Nuclei of C2C12 myotubes (MT) treated with 1.5% 1,6-hexanediol (1,6-HD) or 1.5% 2,5-hexanediol (2,5-HD). Left: Representative images of 4,6-diamidino-2-phenylindole (DAPI)-stained nuclei. Scale bar, 5 μm. Middle: Time-course quantification of the number of heterochromatin foci per nucleus. Right: Boxplots show foci area (μm 2 , y -axis) at 5, 10, and 15 min posttreatment ( x -axis). n = 55 nuclei, three biological replicates. ( B ) Representative live-cell images of Hoechst 33342-stained MT nuclei before and after 1,6-HD treatment (0 and 15 min, respectively), taken from . Arrows indicate changes in heterochromatin foci intensity (pink, increased; blue, decreased), and the red arrow highlights an alteration in chromocenter integrity. ( C ) Number of heterochromatin foci per nucleus in MT following recovery from 1.5% 1,6-hexanediol (1,6-HD) treatment for 15 min, measured at indicated time points. n = 68 nuclei, three biological replicates. ( D ) Representative images of nuclei of myoblast (MB) and MT with or without 1,6-HD treatment (1.5%, 15 min). Right: Quantification of the number of heterochromatin foci per nucleus. n = 40 (MB), n = 60 (MT), three biological replicates. ( E ) Quantification of colocalization between indicated proteins and DAPI foci in MT with or without 1,6-HD treatment (1.5%, 15 min), by Pearson’s correlation coefficients. n = 60, three biological replicates. ( F ) Boxplot showing the distribution of Z -score of the interchromosomal interaction frequencies in MB and MT. ( G ) RNA FISH using ChRO1 and LacZ biotinylated probes. Biotin signal was detected by Fluorescein-conjugated Avidin DCS and amplified with biotinylated anti-Avidin and additional Fluorescein Avidin DCS. Right: Quantification of colocalization between biotin signal and DAPI foci. n = 50 nuclei. ( H ) Number of heterochromatin foci per nucleus of mouse fibroblast cells (NIH3T3) with or without doxycyline (Dox)-induced ChRO1a expression and/or 1,6-HD treatment (1.5%, 15 min). (EV; empty vector). n = 50, three biological replicates. ( I ) Number of heterochromatin foci per nucleus in MB with or without Dox-induced ChRO1a fragment (1–413, CUR) expression and/or 1,6-HD treatment (1.5%, 15 min). n = 75, three biological replicates. Statistical analyses and data presentation details are described in the “Materials and methods” section.

Article Snippet: C2C12 murine myoblast cells and NIH3T3 mouse fibroblast cells were obtained from the American-type culture collection and grown in a growth medium (GM) consisting of Dulbecco’s modified Eagle medium (DMEM) with 10% (v/v) fetal bovine serum at 37°C and 5% CO 2 .

Techniques: Staining, Avidin-Biotin Assay, Amplification, Expressing, Plasmid Preparation