Research Paper Volume 14, Issue 20 pp 8221—8242

Figure 2. Knock down of Rubicon suppresses drug-induced autophagosome formation but does not appear to alter autophagic flux. HCN2 neuronal cells and BV2 microglial cells were transfected with a scrambled siRNA or with an siRNA to knock down the expression of Rubicon and were co-transfected with a plasmid to express LC3-GFP-RFP. After 24h, cells were treated with vehicle control, AR12 (2 μM), neratinib (50 nM) or the drugs in combination for 4h and 8h. The mean number of intense GFP+RFP+ and RFP+ punctae per cell was determined (n = 3 +/-SD) # p < 0.05 greater than vehicle control; ## p < 0.05 greater than AR12 alone value; † p < 0.05 less than corresponding value in siSCR cells; ∞ p < 0.05 greater than corresponding value at the 4h timepoint.