Journal: PLoS ONE

Article Title: Nicotine Stimulates Nerve Growth Factor in Lung Fibroblasts through an NFκB-Dependent Mechanism

doi: 10.1371/journal.pone.0109602

Figure Lengend Snippet: Using a transwell system, primary murine lung fibroblasts were cultured on an upper chamber transwell membrane and primary murine ASM cells were cultured on the lower plastic chamber. Both chambers were cultured in serum free medium. The upper chamber containing fibroblasts was treated with of nicotine (50 µg/ml) for 72 hours. Primary murine ASM cells were harvested for protein isolation. Phosphorylated (p-MLC) and total myosin light chain (total MLC) expression was measured by using immunoblot analysis with densitometry. ASM cells cultured in proximity to nicotine treated fibroblasts ( N ) have significantly increased higher p-MLC/total MLC ratio compared to controls ( C ). n = 6, *p<0.05, error bars represent ±SEM.

Article Snippet: Human ASM cells were purchased through Lonza (Portsmouth, NH) and cultured according to the manufacturer’s protocol.

Techniques: Cell Culture, Membrane, Isolation, Expressing, Western Blot