Limiting of the Innate Immune Response by SF3A-Dependent Control of MyD88 Alternative mRNA Splicing (original) (raw)
Figure 4
qPCR assay demonstrates that inhibition of SF3A1 or SF3B1 enhances production of MyD88S mRNA.
(A) Depicts the MyD88L and MyD88S alternate mRNA splice forms as well as the location of the primers used to monitor the production of MyD88L or MyD88S. qPCR primers to detect either MyD88L or MyD88S are shown in black. Reverse transcription primers to detect MyD88S are shown in red. Reverse transcription primers to detect both MyD88L and MyD88S simultaneously are shown in green. (B) The indicated siRNAs were transfected into RAW264.7 cells, the cells were stimulated with LPS as indicated, and MyD88L and MyD88S mRNA levels were monitored by qPCR. (C) Cells were transfected with the indicated siRNAs, were stimulated with LPS, and MyD88L and MyD88S mRNA levels were monitored by qPCR. (D) RAW264.7 cells were treated with 1.1 ng/ml SSA for six hours, were subsequently exposed to LPS for six hours (in the presence of SSA), and βactin, MyD88L and MyD88S mRNA levels were monitored by qPCR (normalized relative to βactin primers that cross intron 3). RNA of 1 is defined as MyD88L or MyD88S mRNA levels in the presence of the control non-targetting siRNA in the absence of LPS (B) or in the presence of LPS (C and D).