Journal: Cell reports

Article Title: De novo assembly of RNA m 6 A modification factors into viral genome-associated nuclear bodies drives HCMV RNA accumulation

doi: 10.1016/j.celrep.2025.115826

Figure Lengend Snippet: (A and B) NHDFs infected with HCMV TB40/E in the presence of CHX were fixed at 6 hpi. YTHDC1 was visualized by indirect IF, and HCMV IE1/2 mRNA (A) or HCMV RNA5.0 (B) was visualized using smFISH probes. Scale bar, 10 μm. (C and D) As in (A), except infected cells were identified using indirect IF for HCMV IE1/2 proteins, and the cellular GAPDH (C) or IFNB1 (D) mRNA was visualized using smFISH probes. (E) HCMV TB40/E-infected cells were permeabilized at 6 hpi and subsequently treated with PBS or RNase A before fixation and YTHDC1 or WTAP visualization. DAPI stained nuclei are included in the merged images. Scale bar, 20 μm.

Article Snippet: Human GAPDH FISH probes , Molecular Instruments , RTD640.

Techniques: Infection, Staining

Journal: iScience

Article Title: β-adrenergic signaling modulates breast cancer cell mechanical behaviors through a RhoA-ROCK-myosin II axis

doi: 10.1016/j.isci.2025.112676

Figure Lengend Snippet: βAR activation results in increased cellular force generation (A) Schematic illustration showing the traction force microscopy (TFM) assays. (i) TFM-Pillars Assay. Gold disks (yellow) are embedded on top of polydimethylsiloxane (PDMS) pillars to facilitate imaging lateral displacements due to cellular traction forces. (ii) TFM-Beads Assay. Gold nanoparticles (yellow) are embedded in a PDMS matrix to facilitate imaging lateral displacements due to cellular traction forces. (B) Representative images of MDA-MB-231 HM cells on micropillars with superimposed vector force map. Color scale indicates the force per pillar. Scale, 4 μm. (C) Traction forces of MDA-MB-231 HM cells using TFM-Pillars assay after treatment for 24 h with: vehicle, Veh; βAR agonist isoproterenol, Iso (100 nM); βAR antagonist propranolol, Pro (10 μM); or 100 nM Iso and 10 μM Pro. Each dot represents an individual pillar from at least 16 single cells across 3 independent experiments. Bars show the median; error bars represent standard error. (D) Representative images from traction force measurements of TNBC and MCF10A cells treated with isoproterenol (Iso) for 24 h using the TFM-Beads assay. Each arrow indicates the direction of force and the color gradient corresponds to the magnitude of stress. Scale: 10 μm. (E) Quantification of traction forces. Each data point represents the average traction force per bead for an individual cell, which is averaged over multiple beads that are within the boundary of each individual cell. Data shown here represent at least 7 individual cells across 3 independent experiments. Bars show the median; error bars represent standard error. (F) Western blotting against mono-phosphorylated myosin light chain 2 (pMLC2), non-phosphorylated MLC2, and GAPDH from MDA-MB-231 HM cells treated with 100 nM of isoproterenol for up to 48 h. Quantification of band intensity for pMLC2/MLC2 was normalized to 0 h sample. (G) Western blotting against di-phosphorylated MLC2 (ppMLC2), non-phosphorylated MLC2, and GAPDH after MDA-MB-231 HM cells were treated with increasing concentrations of isoproterenol for 2 h. The ratio of ppMLC2 to MLC2 was normalized to vehicle control. (H) Expression levels of ADRB2 transcripts in TNBC, MCF10A, and MCF7 cells measured by qRT-PCR. Images in (A) are adapted from Servier Medical Art by Servier and are published a Creative Commons BY license ( https://creativecommons.org/licenses/by-nc/3.0/ ). Unless otherwise stated, all error bars represent mean ± s.e.m ( N = 3). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001 [one-way ANOVA with Tukey’s test (F–H) and statistical significance in (C and E) is determined using a permutation test to evaluate the difference in medians between control and treatment conditions. ∗ p < 0.05].

Article Snippet: Primer-probes for human ADRB2 , ThermoFisher , Hs00240532_s1.

Techniques: Activation Assay, Microscopy, Imaging, Plasmid Preparation, Western Blot, Control, Expressing, Quantitative RT-PCR