Fig. 5

RELA KO is reproducible in a wide variety of primary chondrocyte populations and in a chondrocyte cell line with comparably high editing efficiency. (A) Editing efficiency and (B) cellular viability data of OA chondrocytes, human FE002 primary chondroprogenitors, the C28/I2 cell line and bovine chondrocytes edited with sgRNA #1 to induce a RELA KO. Cells mock-transfected with an incomplete RNP (i.e., Cas9 only, no sgRNA) were used as a control. RT-qPCR of selected inflammatory pathways acting downstream of NF-κB in (C) human and (D) bovine WT and KO cells following stimulation with IL-1β for 16 h. Data are represented as mean ± standard deviation of 3 technical replicates for bovine chondrocytes (n = 3) and the C28/I2 cell line (n = 3), of 3 technical replicates from two primary chondroprogenitor donors (n = 6), and of 3 technical replicates from three OA chondrocyte donors (n = 9). Statistical significance was determined using an unpaired t-test (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001)