Western Blot Analysis of Signaling Pathways

Western blot was performed using TF1-BA and TF1-BAP cells treated with IM (2 M), AG490 (25 M) or E6201 (100 nM) during 24 hours (h), proteins were then extracted using RIPA buffer and 40 g of proteins were loaded. Membranes were incubated with monoclonal-antibodies against P-ERK (#4370), ERK (#4695), P-STAT3 (#9145), STAT3 (#9139), Caspase 3 (#9662). P-ABL (#2865), P-CRKL (#3181), P-Smad1/5/8 (#13820), GAPDH (#8884) (Cell Signalling Technology), BCR-ABL (ThermoFisher, #MA1-153) and Smad (SantaCruz Technology, #sc6031).^{21 (link)}

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Jeanpierre S., Arizkane K., Thongjuea S., Grockowiak E., Geistlich K., Barral L., Voeltzel T., Guillemin A., Gonin-Giraud S., Gandrillon O., Nicolini F.E., Mead A.J., Maguer-Satta V, & Lefort S. (2020). The quiescent fraction of chronic myeloid leukemic stem cells depends on BMPR1B, Stat3 and BMP4-niche signals to persist in patients in remission. Haematologica, 106(1), 111-122.

Publication 2020

P-CRKL P-Smad1/5/8 GAPDH BCR-ABL

Ag490 Buffer Caspase 3 Cells Crkl Gapdh Membranes Monoclonal antibodies Proteins Proteins g Ripa Stat3 Western blot

Corresponding Organization :

Other organizations : Centre de Recherche en Cancérologie de Lyon, Institute of Molecular Medicine, École Normale Supérieure d'Abidjan, École Normale Supérieure, Laboratoire de Biologie Moléculaire et Cellulaire des Eucaryotes, Université Claude Bernard Lyon 1

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Variable analysis

independent variables

IM (2 μM)
AG490 (25 μM)
E6201 (100 nM)

dependent variables

Protein expression of P-ERK, ERK, P-STAT3, STAT3, Caspase 3, P-ABL, P-CRKL, P-Smad1/5/8, GAPDH, BCR-ABL, Smad

control variables

TF1-BA and TF1-BAP cell lines
Treatment duration of 24 hours
Protein extraction using RIPA buffer
Loading of 40 μg of proteins

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