Protein Level Changes in Wound Healing Cells

AMG-treated fibroblasts as well as unwounded and MG-treated keratinocytes were used to evaluate changes in protein levels upon treatment (untreated cells were used as controls). Samples were processed as [14 (link)]. Membranes were then incubated overnight at 4 °C with primary antibodies: rabbit α-Smooth Muscle actin (1:800, ab15734, Abcam), mouse p-ERK (1:1000, sc-7383, Santa Cruz), rabbit ERK (1:1000, sc-94, Santa Cruz), mouse Tubulin-alpha (1:1000, T5168, Sigma Aldrich). Analyses were performed using the ChemiDoc XRS + detection system (BioRad, Temse, Belgium) in combination with chemiluminescent HRP substrates SuperSignal West Pico PLUS and SuperSignal West Femto (Thermo Fisher Scientific). Relative densitometry was obtained normalizing all samples to the housekeeping Tubulin-alpha, using both the QuantityOne software and ImageLab software (BioRad).

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Balli M., Vitali F., Janiszewski A., Caluwé E., Cortés-Calabuig A., Carpentier S., Duelen R., Ronzoni F., Marcelis L., Bosisio F.M., Bellazzi R., Luttun A., De Angelis M.G., Ceccarelli G., Lluis F, & Sampaolesi M. (2019). Autologous micrograft accelerates endogenous wound healing response through ERK-induced cell migration. Cell Death and Differentiation, 27(5), 1520-1538.

Publication 2019

ab15734 mouse p-ERK sc-94 ChemiDoc XRS + detection system SuperSignal West Pico PLUS SuperSignal West Femto QuantityOne software ImageLab software

Actin Antibodies Cells Densitometry Erk 1 Fibroblasts Keratinocytes Membranes Mouse Protein changes Rabbit Smooth muscle Tubulin alpha

Corresponding Organization :

Other organizations : KU Leuven, University of Arizona, University of Pavia

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Variable analysis

independent variables

AMG treatment
MG treatment

dependent variables

Protein levels

control variables

Untreated cells

controls

Positive control: Untreated cells
Negative control: Not explicitly mentioned

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