Figure 8. Silencing of tm-chemokine expression abolishes the s-chemokine mediated activation in responsive glioma cell lines and primary cultures.
Cell lines and primary cells were transfected with CXCL16 or CX3CL1 specific RNAi (or non-specific control RNAi), left for recovery for 36 hr and stimulated with 1nM CXCL16 or CX3CL1 for 15 min. Samples were analyzed by SDS-PAGE separation and immunoblotting for phosphorylated ERK 1/2, and re-probed for ERK 2 to ensure equal loading. Silencing efficiency and basic transcription level were confirmed by quantitative RT-PCR (right panel). In tm-chemokine silenced cultures, no activation can be observed by incubation with the respective s-chemokine. Experiments with T98G were repeated in 2 independent biological replicates, shown primary cultures are representative examples from 2 different patient’s primary cultures (compare Figure 8—figure supplement 1).
Figure 8—figure supplement 1. Biological replicates of western blot experiments from siRNA knockdown in T98G glioma cells (compare Figure 8).
