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. Author manuscript; available in PMC: 2021 Dec 1.
Published in final edited form as: Ann Rheum Dis. 2021 Jul 5;80(12):1615–1627. doi: 10.1136/annrheumdis-2021-220000

Fig. 3. 14-3-3ε is required for PGRN regulation of chondrocyte metabolism.

Fig. 3.

(a) Schematic for generating 14-3-3ε−/− human C28I2 chondrocytes using CRISPR/Cas9 technology. (b) Western blotting to confirm the loss of 14-3-3ε in 14-3-3ε knockout C28I2 cells. Cell lysates were examined by immunoblotting with 14-3-3ε antibody. (c) mRNA levels of Col2, Acan and COMP in control and 14-3-3ε knockout C28I2 cells treated with or without 200ng/ml PGRN for 24hrs, assayed by qRT-PCR analysis. (d) mRNA levels of Mmp13, Adamts5, Cox2 and Nos2 in control and 14-3-3ε knockout C28I2 cells treated with 10ng/ml TNFα in the absence or presence of 200ng/ml PGRN for 24 hrs, assayed by qRT-PCR analysis. (e) Expression of Flag-14-3-3ε in control and 14-3-3ε knockout C28I2 cells, assayed by western blot. (f) mRNA levels of Col2 and Acan in PGRN (200ng/ml) treated control or 14-3-3ε knockout C28I2 cells with or without re-expression of 14-3-3ε, assayed by qRT-PCR analysis. (g) Control and 14-3-3ε knockout C28I2 cells with or without re-expression of 14-3-3ε were treated with 10ng/ml TNFα in the absence or presence of 200ng/ml PGRN for 24 hrs. mRNA levels of Mmp13, Adamts5, Cox2 and Nos2 were measured by qRT-PCR. Data are mean ± SD; n = 4 biological replicates; * P < 0.05 or ** P < 0.01.