Concatemeric Broccoli reduces mRNA stability and induces aggregates
Fig 1
Broccoli tandem array synthesis and expression.
(A) Cloning schematic of the constructs containing different copy numbers of Broccoli. (B, C, D) 293T cells were transfected with mCherry or 4x, 8x, 16x, 32x, 64x or 128x Broccoli and analyzed 24 hours post-transfection by flow cytometry. Percentage of Broccoli and mCherry positive cells shown in dot plots (B) and mean fluorescence intensity of Broccoli (C) and mCherry (D) compared between constructs. (D) The mean fluorescence intensity (MFI) of Broccoli in the graph was normalized to the 4x Broccoli construct and the MFI of the remaining constructs was calculated as follows: ⦗ (MFI XxBroccoli): (4xBroccoli) ⦘. Data representative of three independent experiments. Statistical analysis performed by t-test (c, d). * P<0.05, ** P<0.01, *** P<0.001.