Ab76464

Total protein was extracted by RIPA lysis buffer (Keygen). After detection of protein concentration, extracted protein was separated with SDS‐PAGE (Beyotime). After transferring onto PVDF membranes (Millipore), the membranes were blocked in non‐fat dried milk (Yili). After that, the membranes were probed with unique primary antibodies: KI67 (ab181255; 1:5000; Abcam), Bax (ab32503; 1:3000; Abcam), multidrug resistance protein (MRP1; ab76464; 1:1000; Abcam), CRABP2 (ab181255; 1:3000; Abcam), and β‐actin (ab8226; 1:1000; Abcam), and then secondary antibody was used for combination with primary antibodies. At last, the combined signals were detected by ECL reagent (Abcam).

Extraction of total protein of A549 and H1299 cells was performed with RIPA lysis buffer (Beyotime, Shanghai, China), protein concentration was determined by BCA method, and then the protein samples were mixed with loading buffer and denatured in boiling water. Subsequently, 15 μL of protein sample in each group was loaded into each well and the protein was dissolved by SDS-PAGE. After the protein was transferred to PVDF membrane (Millipore, Bedford, MA, USA), the PVDF membrane was blocked with 5% skimmed milk at room temperature for 1 h. Then the primary antibody (rabbit anti-FGFR1 monoclonal antibody, ab76464, Abcam, Shanghai, China, 1:1000) was used to incubate the PVDF membrane at 4°C overnight and then rinsed with TBST 3 times with 5 min each time. The corresponding secondary antibody (goat anti-rabbit IgG, ab205718, Abcam, Shanghai, China. 1:2000) was then applied to incubate the membrane at room temperature for 1.5 h before the membrane. After that, the membrane was rinsed with TBST 3 times with 5 min each time. Next, the protein band was developed employing ECL kit (Beyotime Biotechnology, Shanghai, China), and the gray values of protein bands were quantitatively analyzed utilizing ImageJ (National Institutions of Health, Bethesda, MD, USA). GAPDH was used as the internal reference.