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. 2017 Jan 5;7:663. doi: 10.3389/fimmu.2016.00663

Figure 6.

Figure 6

Cathepsin B activity is required for an optimal response to QS-21. (A) moDCs were incubated with CA-074 Me (cathepsin B inhibitor), Z-FA-FMK (cathepsin B inhibitor), Z-FF-FMK (cathepsin B/L inhibitor), or pepstatin A (cathepsin D, cathepsin E, and pepsin inhibitor) for 1 h and stimulated with QS-21 for 24 h. TNF release into the supernatant was measured by ELISA. Statistical significance was determined with a Friedman test followed by Dunn’s multiple comparison test. (B) moDCs were pretreated with CA-074 Me or Z-FA-FMK for 1 h and stimulated with QS-21 or MPL for 6 h. IL-6 and TNF mRNA levels were quantified by qPCR and normalized to β-actin mRNA levels. Statistical significance was determined by two-way ANOVA followed by Tukey’s multiple comparisons test. (C,D) moDCs were transduced with lentiviruses coding for shRNAs targeting GAPDH (shGAPDH) or cathepsin B (shCtsb), and cathepsin B (C), IL-6 and TNF (D) mRNA expression were measured by qPCR and normalized against β-actin expression. Statistical significance was determined by a Wilcoxon matched-pairs signed-rank test.