Figure 6 .

COL1A1 expression in cultured donor stromal fibroblasts after siRNA knockdown of (A) CENPF and HSPG2, and (B) EML6 and NBEAL2. Cells were electroporated with 100 nM SMARTpool siRNAs (Dharmacon) for each targeted gene individually, and COL1A1 expression, measured as a functional readout of stromal cells. Inhibition of CENPF (95%), HSPG2 (44%), EML6 (84%) and NBEAL2 (60%). Only knockdown of CENPF and HSPG2 resulted in 20–51% increase in COL1A1 expression, respectively. Gene expression was measured by Taqman qPCR and relative expression normalized to 18S RNA determined using the 2^-DDCt (cycle threshold) method. Error bars represent SEs of the mean for 3 replicates and all pairwise comparisons are with transfections using control siRNAs. (^*P ≤ 0.05, ^**P ≤ 0.01, ^***P ≤ 0.001, ^****P ≤ 0.0001, ns—not significant).