Ab238825

IHC staining was performed to evaluated the expression of MCT1 and MCT4, in both tumors and paired adjacent lung tissues in each case. In brief, the formalin-fixed paraffin-embedded tissues were cut into a series of 5 µm-thick sections. The sections were de-paraffinized, rehydrated and underwent antigen retrieval using Dako EnVisionTM FLEX Target Retrieval Solution (pH 9.0) at 95 °C for 20 minutes. Anti-SLC16A3 (MCT4) antibody (Rabbit polyclonal antibody, HPA021451, Sigma-Aldrich), and anti-MCT1 antibody (Rabbit polyclonal antibody, ab238825, Abcam) were used as primary antibodies, and were diluted at a ratio of 1:100. After 2 hours of primary antibody incubation at room temperature, incubation with a secondary antibody (PV-9000, Zhongshan Jinqiao Biotechnology Co. Ltd., Beijing, China) was performed at room temperature for 30 minutes. Finally, 3,3’-diaminobenzamine and hematoxylin were used for coloration of the immune complex and nucleus, respectively. The expression of MCT4 and MCT1 was assessed by multiplying the staining intensity score and the percentage score as described in our previous study (26 ). A final score ≥6 was defined as high expression.

Cell lines or tissues were centrifuged at 14,000 g for 15 min at 4 °C after being extracts with RIPA lysis buffer. SDS-PAGE was used to separate the proteins and PVDF membrane was used to transfer them. In this investigation, the following primary antibodies were utilized: MYBL2 (ab238825, Abcam, 1:1000), SMO (ab52771, Abcam, 1:1000), Bax (ab32503, Abcam, 1:1000), Bcl-xl (ab32124, Abcam, 1:1000), and GAPDH (ab8245, Abcam, 1:1000); Protein signals were detected using a chemiluminescence kit from Bio-Rad.