Antibodies against stat1

PLSCR1 shRNA and STAT3 shRNA were purchased from Sigma-Aldrich (St Louis, MO). Human PLSCR1, STAT3, and EGFR were amplified from the MDA-MB231 cDNA and subcloned into pLenti6.3⁄V5, pLVX, and pCMV, respectively.
Recombinant human EGF protein, human interleukin-6 (hIL-6), and antibodies against PLSCR1 and ALDH1 were purchased from Abcam (Carlsbad, CA). Antibodies against STAT1, STAT3, Phospho- STAT1 (Tyr701) and Phospho-STAT3 (Tyr705) were acquired from Cell Signaling Technology (Danvers, MA). Antibodies against phospho-Tyr were obtained from Santa Cruz (Dallas, TX). 6x-His Tag monoclonal antibody and Alexa Fluor 555-conjugated goat anti-rabbit IgG were purchased from Thermo Fisher Scientific. Antibodies for Flag, Myc, β-Tubulin, LaminB and Anti-FLAG Magnetic Beads were obtained from Sigma-Aldrich (St. Louis, MO).

Dendritic cells were collected and lysed with lysing buffer (Sigma). Protein extracts were separated on 10% polyacrylamide-SDS gels and electroblotted onto nitrocellulose membranes (Gene script). After blocking with 5% nonfat dry milk in TBS, the membranes were incubated with antibodies against STAT1, P38 MAPK, phosphorylated STAT1, and phosphorylated p38 MAPK (Cell Signaling Technology), followed by incubation with HRP-conjugated secondary antibody (Cell Signaling Technology). The blots were normalized to GAPDH.