Immunoblotting Analysis of Cellular Proteins

Cell lysates were prepared and immunoblotting was carried out as previously described.^{13 (link)} Anti-TYRO3 (Ref. 5585), AXL (Ref. 8661), MERTK (Ref. 4319), caspase-8 (Ref. 9746), cleaved caspase-3 (Ref. 9664), cleaved PARP (Ref. 5625), FOXM1 (Ref. 5436), pRB (Ref. 9309), phospho-pRB Ser780 (Ref. 9307), CYCLIN D1 (Ref. 2926), AURORA A (Ref. 14475), AURORA B (Ref. 3094), SURVIVIN (Ref. 2808), c-MYC (Ref. 9402), anti-mouse IgG, HRP-linked (Ref. 7076) and anti-rabbit IgG, HRP-linked (Ref. 7074) antibodies were purchased from Cell Signaling Technology (Ozyme, Montigny-le-Bretonneux, France). Anti-α-TUBULIN (Ref. T6199) and anti-β-ACTIN (Ref. A2228) antibodies were obtained from Sigma-Aldrich. For peptide-N-glycosidase F (PNGaseF), we digested 10 µg of protein with PNGaseF, according to the manufacturer’s instructions (New England Biolabs, Evry, France).

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Dufour F., Silina L., Neyret-Kahn H., Moreno-Vega A., Krucker C., Karboul N., Dorland-Galliot M., Maillé P., Chapeaublanc E., Allory Y., Stransky N., Haegel H., Menguy T., Duong V., Radvanyi F, & Bernard-Pierrot I. (2019). TYRO3 as a molecular target for growth inhibition and apoptosis induction in bladder cancer. British Journal of Cancer, 120(5), 555-564.

Publication 2019

Anti-TYRO3 MERTK caspase-8 cleaved caspase-3 cleaved PARP FOXM1 phospho-pRB Ser780 CYCLIN D1 AURORA A AURORA B SURVIVIN c-MYC anti-mouse IgG, HRP-linked Anti-α-TUBULIN anti-β-ACTIN PNGaseF

Actin Anti igg Antibodies Aurora a Aurora b C myc Caspase 3 Caspase 8 Cell Cyclin d1 Mertk Mouse Peptide n glycosidase f Protein Rabbit Survivin α tubulin

Corresponding Organization :

Other organizations : Centre National de la Recherche Scientifique, Institut Curie, Université Paris Sciences et Lettres, Assistance Publique – Hôpitaux de Paris, Hôpitaux Universitaires Henri-Mondor, Université Paris Cité, Sorbonne Université, Cherry Biotech (France)

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Variable analysis

independent variables

Cell lysates were prepared and immunoblotting was carried out as previously described.

dependent variables

Protein expression levels of TYRO3, AXL, MERTK, caspase-8, cleaved caspase-3, cleaved PARP, FOXM1, pRB, phospho-pRB Ser780, CYCLIN D1, AURORA A, AURORA B, SURVIVIN, and c-MYC

control variables

Anti-α-TUBULIN and anti-β-ACTIN antibodies were used as loading controls

controls

Positive control: Digestion of 10 µg of protein with PNGaseF according to the manufacturer's instructions
Negative control: Not explicitly mentioned

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