Journal: Scientific Reports

Article Title: The interaction of mast cells with membranes from lung cancer cells induces the release of extracellular vesicles with a unique miRNA signature

doi: 10.1038/s41598-023-48435-4

Figure Lengend Snippet: The phosphorylation of ERK 1/2 in HMC-1 cells stimulated by H1299-derived membranes was evaluated upon inhibition of MEK, PI3K, PKC, A3R, and CD73. HMC-1 cells (1 × 10 6 cells/ml) were pretreated with the indicated inhibitors (3 µM U0126, 10 µM LY 294002, 1 µM GO 9676, 100 nM MRS 1220, and 5 µM APCP, respectively) for 30 min. Subsequently, the cells were exposed to 10 µg/ml H1299-derived membranes for 1 min. Cellular lysates underwent SDS-PAGE separation and were subjected to probing with an anti-p-ERK 1/2 antibody and subsequent reprobed using an anti-total ERK2 antibody. The phospho-ERK 1/2 and total ERK bands were determined using ImageJ software, and the relative pixel densities (phosphorylated/total) were calculated and normalized to those of untreated (UT) cells. The data presented reflect the average of three distinct experiments and are shown as the mean ± SEM (error bars). A p value less than 0.05 was considered significant and denoted by *, while ** indicates a p value less than 0.01.

Article Snippet: The PKC inhibitor GO6976, PI3K inhibitor LY294002, MEK inhibitor U0126, and CD73 inhibitor APCP were purchased from A.G. Scientific Inc.

Techniques: Derivative Assay, Inhibition, SDS Page, Software