Sall4

The cells were lysed in RIPA buffer containing proteinase inhibitors. Equal amount of proteins was loaded and separated on a 12% SDS-PAGE gel. Following electrophoresis, the proteins were transferred to a PVDF (polyvinylidene difluoride) membrane, blocked in 5% (w/v) non-fat milk, and incubated with the primary antibodies at 4 ^oC overnight. The sources of primary antibodies were: SALL4 (954-1053; Abnova, Taipei, Taiwan); CD44 (BS6825; Bioworld technology, Louis Park, MN, USA) and GAPDH (MB001, Bioworld technology); ERK (4695S; Cell Signaling Technology, Beverly, MA, USA), p-ERK (4370S; Cell Signaling Technology), NF-κB p65 (8242S; Cell Signaling Technology), p-NF-κB p65 (3033P; Cell Signaling Technology), STAT3 (4904P; Cell Signaling Technology), p-STAT3 (9145P; Cell Signaling Technology), N-cadherin (13116S; Cell Signaling Technology), and Oct4 (2750S; Cell Signaling Technology); Sox2 (AB5603; Merck Millipore, Shanghai, China), c-Myc (10057-1-AP; Proteintech, Rosemont, IL, USA), Nanog (AF1505; Signalway Antibody, College Park, MD, USA), E-cadherin (H-108, Santa Cruz Biotechnology, Dallas, TX, USA). After washing with TBS/T for three times, the membranes were incubated with HRP-conjugated goat anti-rabbit or anti-mouse secondary antibodies (Bioworld technology) at room temperature for 1 h. The protein bands were visualized by enhanced chemiluminescence.

Cells were harvested and lysed in RIPA supplemented with protease inhibitors. Equal amounts of protein lysates were separated by SDS-PAGE and electrically transferred to nitrocellulose membrane. The membranes were probed with the following specific primary antibodies: SALL4 (Abnova, H00057167-M03), E-cadherin (Cell Signaling Technology, #3195), vimentin (Cell Signaling Technology, #5741), Oct4 (Abcam, ab18976), Bmi-1 (Abcam, ab126783), Nanog (Cell Signaling Technology, #4903), NF-κB p65 (Cell Signaling Technology, #8242), Phospho-NF-κB p65 (Cell Signaling Technology, #3033), Sox2 (Cell Signaling Technology, #14962), and β-actin (Cell Signaling Technology, #12262). Then they were blotted with an HRP (horseradish peroxidase)-conjugated secondary antibody. Blots were visualized by enhanced chemiluminescence (ECL).