Irdye700 800 mouse or rabbit

Total protein was collected from cells with RIPA lysis Mix. Western blotting assay was performed as previously described [23 (link)]. Briefly, 60 μg protein extractions were loaded via SDS-PAGE and transferred onto nitrocellulose membranes (absin, China), then incubated with primary antibodies for 2 hrs at temperature, then plated at 4 °C overnight, the membranes were incubated in 5% non-fat milk blocking buffer for horizontal mode 3 h. After incubation with secondary antibodies IRDye700/800 Mouse or Rabbit (Lincoln, Nebraska, USA), the membranes were scanned using an Odyssey, and data were analyzed with Odyssey software (LI-COR, USA). Primary antibodies list: DDR1 (SAB1300850, Sigma, USA), Vimentin (10366-1-AP, Proteintech, USA), N-cadherin (22018-1-AP, Proteintech, USA), E-cadherin (20874-1-AP, Proteintech, USA), GLUD1 (14299-1-AP, Proteintech, USA), GLS1 (12855-1-AP, Proteintech, USA), SLC1A5 (20350-1-AP, Proteintech, USA), STAT3 (10253-2-AP, Proteintech, USA), p-STAT3(705) (ab76315, Abcam, UK), p-STAT3(727) (ab32143, Abcam, UK), GAPDH (60004-1-Ig, Proteintech, USA).

After RIPA cleavage, we extracted total protein and measured it with BCA method. After quantitative denaturation, proteins were separated using 10 or 15% polyacrylamide gels and transferred onto 0.22 μm PVDF membranes (Merck Millipore, United States). The first incubation and second incubation were carried out according to the operation steps. The expression of the protein was expressed by the gray value. Primary antibodies list: GADPH (ab181602, Abcam), cleaved-caspase3 (ab2302, Abcam), Bax (ab32503, Abcam), Bcl2 (12789-1-AP, Proteintech), TNFα (17590-1-AP, Proteintech), and IL-6 (66146-1-Ig, Proteintech). The secondary antibodies IRDye700/800 Mouse or Rabbit were produced by LICOR (Lincoln, Nebraska, United States) for 1 h, and the bands were scanned using Odyssey.

After RIPA cleavage, we extracted total protein and measured with BCA method. After quantitative denaturation, protein electrophoresis membrane transfer and blocked. The first incubation and second incubation were carried out according to the operation steps. The expression of the protein was expressed by the gray value. Primary antibodies list: GADPH (ab181602, Abcam), cleaved-caspase3 (ab2302, Abcam), bax (ab32503, Abcam), bcl2 (12789-1-AP, Proteintech), TNFα (17590-1-AP, Proteintech), IL 6 (66146-1-Ig, Proteintech), p-IκBα ((ab92700, Abcam), IκBα (10268-1-AP, Proteintech), p65 (66535-1-Ig, Proteintech) and p-p65 (ab183559, Abcam). The secondary antibodies IRDye700/800 Mouse or Rabbit were produced by LICOR (Lincoln, Nebraska, USA).