Rabbit monoclonal anti phosphorylated stat3

For immunofluorescence, cells were washed with PBS and fixed in 4% paraformaldehyde at room temperature for 30 min. Non-specific binding was blocked using 10% normal goat serum (Sigma). Cells were incubated with the following primary antibodies after being diluted in PBS with 1% bovine serum albumin at 4 °C overnight: rabbit monoclonal anti-phosphorylated Caveolin-1, rabbit monoclonal anti-phosphorylated STAT3, rabbit monoclonal anti- Caveolin-1 (Cell Signaling Technology). Then, cells were washed twice with PBS and incubated with secondary antibodies at 37 °C for 30 min as follows: FITC-conjugated goat-anti-rabbit IgG (Abcam, Cambridge, UK) or TRITC-conjugated goat-anti-rabbit IgG (Sigma, St. Louis, MO, USA). The slides were mounted in mounting medium with 4’, 6-diamidino-2-phenylindole (DAPI; Vector Laboratories, Burlingame, CA, USA) and viewed with a live cell station (Delta Vision, API).

Cells grown and treated as indicated were collected and total protein was extracted. The following primary antibodies were used: rabbit monoclonal anti-α-actin (Abcam), monoclonal anti-COL1A1 (Millipore), rabbit monoclonal anti-phosphorylated SMAD2, rabbit monoclonal anti-phosphorylated SMAD3, rabbit monoclonal anti- SMAD2/3, rabbit monoclonal anti-ERK1/2, or rabbit monoclonal anti-phosphorylated ERK1/2, rabbit monoclonal anti-Akt, rabbit monoclonal anti-phosphorylated Akt, rabbit monoclonal anti-STAT3, or rabbit monoclonal anti-phosphorylated STAT3 (all from Cell Signaling Technology, Inc.). Horseradish peroxidase-conjugated goat-anti-rabbit antibody (Thermo Scientific) was used as a secondary antibody. The control for equal protein loading was assessed with an anti-β-actin antibody (Cell Signaling Technology, Inc.).

Cells grown and treated as indicated were collected and total protein was extracted. The following primary antibodies were used: rabbit monoclonal anti-phosphorylated Caveolin-1, rabbit monoclonal anti-phosphorylated STAT3, rabbit monoclonal anti- Caveolin-1, rabbit monoclonal anti-STAT3 (all from Cell Signaling Technology, Danvers, MA, USA). Horseradish peroxidase-conjugated goat-anti-rabbit antibody (Thermo Scientific, Waltham, MA, USA) was used as a secondary antibody. The control for equal protein loading was assessed with an anti-β-actin antibody (Cell Signaling Technology).