Journal: Journal of Cell Science

Article Title: Molecular insights into profilin 1-dependent regulation of cellular phosphatidylinositol (4,5)-bisphosphate

doi: 10.1242/jcs.265025

Figure Lengend Snippet: PIP 2 attenuation upon loss of Pfn1 is reversible by PLC inhibition. (A,B) Representative PIP 2 staining images (A; 40×) of control and Pfn1 KO MDA 231 cells, treated with either inactive ( U73343 ) or active (U73122) pan-PLC inhibitor drug (magnified images of PIP 2 staining of selected regions of interest are shown alongside). (B) The associated PM PIP 2 quantification for the various groups (all data normalized to the average readout of control cells treated with the U73343 compound). A total of ∼450 cells/group were analyzed from three independent experiments. Scale bar: 40 μm. (C–E) Representative PLCβ3 and Pfn1 immunoblots of lysates of MDA-231 cells following transient transfection of the indicated siRNAs (non-targeting control siRNA transfection served as control); GAPDH blot serves as the loading control (C). Representative PIP 2 staining images fluorescence images (40×) of MDA-231 cells transfected with the indicated siRNAs (magnified images of PIP 2 staining of selected regions of interest of each group are shown underneath) (D). Scale bar: 80 μm. (E) Associated quantification to compare the PM PIP 2 intensity between the various groups of cells based on the results of three independent experiments. In B,E, the box represents the data falling within the 25th to 75th percentile range, and a line denoting the median value of the data set. Whiskers represent 1.5× the interquartile range above and below the 25th and 75th percentiles. * P <0.05; *** P <0.001 (one-way ANOVA with Tukey HSD post-hoc test for multiple comparisons).

Article Snippet: In experiments involving pan-PLC inhibitor U73122 and its inactive control U73343 (Medchem Express, # HY108630 ), the dry compounds were diluted in DMSO to a stock concentration of 10 mM and added to cells at a final concentration of 2 μM after 24 h of serum stimulation.

Techniques: Inhibition, Staining, Control, Western Blot, Transfection, Fluorescence

Journal: Microbiology Spectrum

Article Title: D-glucose uptake inhibits bovine alphaherpesvirus 1 post-binding cell process entry via inhibition of PLC-γ1 signaling in a glucose transporter 1-independent manner

doi: 10.1128/spectrum.02456-25

Figure Lengend Snippet: The effects of U73122 and glucose on BoAHV-1 entry process. ( A and B ) MDBK ( A ) and A549 ( B ) cells in six-well plates were subjected to glucose depletion via culturing the cells using glucose-free DMEM for 12 h. The cells were infected with BoAHV-1 (MOI = 1) in the presence of either U73122 or D-glucose alone, or in combination, at the designated concentrations, at 37°C for 30 min. After extensive washes, genomic DNA was extracted, and the viral genome bound to the cells was quantitatively evaluated using qPCR. The data shown are the means of three independent experiments with error bars representing the SDs. The significance was evaluated by Student’s t -test (*, P < 0.05; **, P < 0.01).

Article Snippet: Phospholipase C (PLC) inhibitor U73122 (cat# HY-13419) and GLUT1-specific inhibitor BAY-876 (cat# HY-100017) were ordered from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Infection