Anti rabbit mtor and phospho mtor

Immunofluorescences (IF) and immunoblots (WB) were performed with the following primary antibodies: anti-rabbit S100A4 (1:500-IF, 1:1000-WB, Millipore), anti-rabbit mTOR and phospho-mTOR (1:100-WB, Cell Signaling), anti-mouse SQSTM1/p62 (1:1000-WB, Abcam), anti-rabbit NF-kB and phospho-NF-kB (1:1000-WB, Cell Signaling), anti-rabbit STAT3 (1:1000-WB, Cell Signaling), anti-rabbit phospho-STAT3 (1:2000-WB, Cell Signaling), anti-rabbit α-SMA (1:1000-WB, 1:500-IF, GeneTex), anti-rabbit N-cadherin (1:1000-WB, GeneTex), anti-mouse glial fibrillary acidic protein (GFAP) (1:500-IF, 1:1000-WB, Cell Signaling), anti-rabbit β-III Tubulin (1:500-IF, Cell Signaling), anti-mouse MyoG (1:200-WB, Hybridoma Bank, USA), anti-mouse platelet-derived growth factor receptor β (PDGFR-β) (1:250-WB, 1:500-IF, Santa Cruz), and anti-mouse GAPDH (1:10000-WB, Calbiochem). Secondary immunoblot antibodies for WB were anti-rabbit (1:2500) and anti-mouse (1:5000) IgG peroxidase-conjugated from Bio-Rad Laboratories (Hercules, CA, USA). Secondary fluorescent antibodies for IF were Alexa-Flour 488-Donkey anti-rabbit (1:200) and Cy3-Donkey anti-mouse (1:200) from Jackson ImmunoResearch Laboratories (West Grove, PA, USA). DAPI was used to stain nuclei (1:1000, Thermo Fisher Scientific, Waltham, MA, USA).

The following primary antibodies were used for immunofluorescence (IF) or western blot (WB): anti-rabbit S100A4 (1:500-IF, 1:1000-WB, Millipore, Burlington, MA, USA), anti-mouse glial fibrillary acidic protein (GFAP) (1:1000-IF, Novus Biologicals, Centennial, CO, USA), anti-rat CD68 (1:500-IF, Abd Serotec, Kidlington, UK), anti-rat CD11b (1:500-IF, Abd Serotec), anti-mouse paxillin (1:500-IF, 1:1000-WB, BD-Biosciences, San Jose, CA, USA), anti-mouse gp91^phox (1:1000-WB, BD-Biosciences), anti-rabbit mTOR and phospho-mTOR (1:1000-WB, Cell Signaling, Danvers, MA, USA), anti-rabbit NF-κB and phospho-NF-κB (1:1000-WB, Cell Signaling), anti-GAPDH (1:5000-WB, Millipore). Secondary fluorescent antibodies for IF were: Cy3 Donkey anti-rabbit (1:200), Alexa-Fluor 488 Donkey anti-rabbit (1:200), Cy3 Donkey anti-mouse (1:200), and Cy5 Donkey anti-rat (1:200) from Jackson ImmunoResearch Laboratories (West Grove, PA, USA). Phalloidin (1:200, Sigma Aldrich, Milan, Italy) was used to stain cells’ actin filaments. DAPI (1:1000, Thermo Fisher Scientific, Waltham, MA, USA) was used to stain nuclei. Anti-rabbit and anti-mouse IgG peroxidase-conjugated secondary antibodies (1:2500) were from Bio-Rad Laboratories (Hercules, CA, USA).