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Fig. 3 | Arthritis Research & Therapy

Fig. 3

From: Autophagy induction contributes to the resistance to methotrexate treatment in rheumatoid arthritis fibroblast-like synovial cells through high mobility group box chromosomal protein 1

Fig. 3

Inhibition of autophagy enhanced the proapoptotic effect of methotrexate (MTX) on osteoarthritis fibroblast-like synovial cells (OA-FLS) and rheumatoid arthritis fibroblast-like synovial cells (RA-FLS). Cells were transfected with control or Beclin-1 small interfering RNA (siRNA), followed by 0.1 μM MTX treatment for 48 h. The expression of Beclin-1 and microtubule-associated protein 1 light chain 3 types I and II (LC3-I and LC3-II, respectively) was verified by Western blot analysis (a). The numbers of dead cells were determined by flow cytometry following annexin V/propidium iodide (PI) staining (b). Verification of poly(ADP-ribose) polymerase (PARP) p85 protein expression by Western blot analysis (c). The results are representative of four experiments using cells from different patients. β-actin was used as a loading control in all experiments. Student’s t test was used for the data analysis. Values in (a) are the mean ± standard deviation (SD). *p < 0.05 vs. control siRNA. Values in (b) and (c) are the mean ± SD. *p < 0.05 vs. OA-FLS transfected with control siRNA. # p < 0.05 vs. RA-FLS transfected with Beclin-1 siRNA

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Arthritis Research & Therapy

ISSN: 1478-6362

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