Figure 4.

Leptin-induced human myometrial smooth muscle cells (HM-SMC) proliferation at 6.25 ng·mL⁻¹ is mediated through ERK1/2 activation. (A, B and D) Western blots of p-ERK1/2. Cells were cultured with leptin at 6.25 ng·mL⁻¹ for 0–5 min (A), 10–60 min (B) and 1–8h (D). The blots are representative of five independent experiments. Quantification: 42 kDa p-ERK1/2 bands were digitized and quantified with QuantityOne. Phosphorylated bands were compared with total of bands formed for each condition and presented as mean values ± SEM. *P < 0.05 versus CTL (non-stimulated cells). (C) P-ERK1/2 immunofluorescence. Fixed cells were labelled with anti-P-ERK1/2 antibody, Alexa fluor 488 and with DAPI for nuclear localization. Five representative pictures were taken with an epifluorescence microscope (×400) in random chosen fields for each labelling. Pictures were then analysed and merged using ImageJ software. Quantification: bars represented the mean ± SEM from four independent experiments. *P < 0.05 versus CTL (non-stimulated cells).