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Fig. 4. Phenotypic classes identified from the RNAi screen. (A)
Protocol for RNAi screening in primary cultures. (B-F) Four distinct
classes of muscle phenotype were distinguished based on the staining of Actin
using phalloidin (left panels, and red in right panels), Mhc (middle panels)
and of nuclei with DAPI (blue in right panels). (B) Wild-type control myotubes
treated with dsRNAs targeting lacZ. (C) Class I (treated with
mew dsRNAs). (D) Class II (treated with sls dsRNAs). (E)
Class III (treated with Mlc2 dsRNAs). (F) Class IV (treated with
Actin dsRNAs simultaneously targeting all Actin isoforms, including
Act42A, Act57B, Act5C, Act79B, Act87E and Act88F). Note that
the four phenotypic classes did not result from fewer fusions, as muscles
contained the same number of nuclei as controls. Arrowheads point to the
nuclei. Scale bar: 15 µm.
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