Skip to main content
Fig. 5 | Arthritis Research & Therapy

Fig. 5

From: Intervening upregulated SLC7A5 could mitigate inflammatory mediator by mTOR-P70S6K signal in rheumatoid arthritis synoviocytes

Fig. 5

Activation of mTOR-P70S6K signaling and downstream upregulation of MMP3 and MMP13 expression by SLC7A5 overexpressed in RA FLS. a The protein expression of p-mTOR (n = 4), mTOR, and P70S6K (n = 3) in synovial tissue from OA and RA patients detected by Western blotting. b The protein synthesis pathway activation in FLS. The FLS were treated with 20 ng/mL IL-1β for 4 h and collected for protein isolation, detected by Western blotting. The density of SLC7A5 immune-reactive bands was analyzed by using ACTB expression as a loading control. The phosphorylation protein ratio fold change of mTOR and P70S6K was analyzed by using total protein expression of their own as a control, while the phosphorylation protein ratio fold change of 4EBP1 was analyzed by using ACTB expression as a loading control (n = 3). c The impact of SLC7A5 siRNA on the protein synthesis pathway (mTOR-P70S6K-4EBP1) activation in FLS, detected by Western blotting. The cells were transfected with siNC or siSLC7A5 (si-2) for 24 h and then stimulated with 20 ng/mL IL-1β for another 4 h. The fold change in phosphorylated/non-phosphorylated protein ratios of mTOR and P70S6K was analyzed by using total protein expression of their own as a control, while that of 4EBP1 was analyzed by using ACTB expression as a loading control (n = 3). d The inhibition of MMP3 and MMP13 expression by rapamycin (mTORC1 inhibitor) in RA FLS under IL-1β treatment. The cells were incubated with rapamycin (100 nM) for 8 h and then stimulated with 20 ng/mL IL-1β for another 24 h. The protein levels were detected by Western blotting. The density of MMP3 and MMP13 immune-reactive bands was analyzed by using ACTB expression as a loading control (n = 3) (*p < 0.05)

Back to article page