Fig. 2
Mechanism of SLC7A5 upregulation in fibroblast-like synoviocytes (FLS) under IL-1β treatment. a The protein expression of SLC7A5 in FLS from RA patients treated with different cytokines (IL-1β 20 ng/mL, TNF-α 10 ng/mL, IFN-γ 20 ng/mL, IL-6 20 ng/mL, and IL-17A 10 ng/mL) for 24 h, detected by Western blotting. The density of SLC7A5 immune-reactive bands was analyzed by using ACTB expression as a loading control (n = 3). b The representative active signaling pathways in FLS from RA patients, detected by Western blotting. The cells were treated with 20 ng/mL IL-1β and collected at different time points for protein isolation. c The mRNA levels of SLC7A5 in FLS from RA patients incubated with different inhibitors and stimulated with IL-1β. The cells were firstly treated with NF-κB inhibitor Bay11-7085 (10 μM) or JNK inhibitor SP600125 (10 μM) for 4 h and then treated with 20 ng/mL IL-1β for 24 h. The mRNA expression was detected by RT-qPCR (n = 6). d The protein expression of SLC7A5 in FLS from RA patients incubated with different inhibitors and stimulated with IL-1β. The cells were firstly treated with NF-κB inhibitor Bay11-7085 (10 μM) or JNK inhibitor SP600125 (10 μM) for 4 h and then treated with 20 ng/mL IL-1β for 24 h. The protein expression was detected by Western blotting. The SLC7A5 immune-reactive band density was analyzed by using ACTB expression as a loading control (n = 3). e The protein expression of SLC7A5 in FLS from RA patients incubated with p38 inhibitor and stimulated with IL-1β. The cells were firstly treated with p38 inhibitor SB203580 (10 μM) for 4 h and then treated with 20 ng/mL IL-1β for 24 h. The protein expression was detected by Western blotting. SLC7A5 immune-reactive bands density was analyzed by using ACTB expression as a loading control (n = 3) (*p < 0.05)