Fig. 5

MiR-4303 relieves chondrocyte inflammation via targeting ASPN. Chondrocytes were randomly divided into four groups, control group; chondrocytes were untreated; NC mimic + pc-NC + LPS group, 1 × 10⁵ chondrocytes were treated with 10 μL LPS (100 ng/mL), 100 nM NC mimics were transfected into chondrocytes, and 500 ng pc-NC were transfected into chondrocytes; miR-4303 mimic + pc-NC + LPS group, 1 × 10⁵ chondrocytes were treated with 10 μL LPS (100 ng/ml), 100 nM miR-4303 mimics were transfected into chondrocytes, and 500 ng pc-NC were transfected into chondrocytes; miR-4303 mimic + pc-ASPN + LPS group, 1 × 10⁵ chondrocytes were treated with 10 μL LPS (100 ng/ml), 100 nM miR-4303 mimics were transfected into chondrocytes, and 500 ng pc-ASPN were transfected into chondrocytes. A RT-qPCR and Western blot were performed to assess miR-4303 and ASPN protein expression levels, respectively. B CCK-8 assay was performed to detect chondrocyte viability. C EdU assay was carried to evaluate chondrocyte proliferation. D Western blot was conducted to measure the levels of proliferation-associated proteins, including PCNA and Ki-67 in chondrocytes. E FACs were performed to evaluate the distribution of chondrocytes in each cell cycle. F Western blot was conducted to measure the levels of cell cycle-related proteins, including CyclinA1, CyclinB1, CyclinD2 and p27. G, FACs were conducted to assess chondrocytes apoptosis. H, Western blot was performed to detect the levels of apoptosis-related proteins, including Bax, Bcl-2, Cleaved-casapase3 and Cleaved-casapase9. I-K, ELISA was performed to detect the levels of TNF-α, IL-1βand IL-6 in the supernatant of chondrocytes. Tukey post-test was used for comparison between multiple groups. *P < 0.05, **P < 0.01 versus control group; ^#P < 0.05, ^##P < 0.01 versus NC mimic + pc-NC + LPS group; ^$P < 0.05, ^$$P < 0.01 versus miR4303 mimic + pc-NC + LPS group. Error bars represented SD. Data represented three independent experiments