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. 2020 Mar 11;12(3):652. doi: 10.3390/cancers12030652

Figure 3.

Figure 3

Inserting the eight amino acids in the HEAT H3 repeat of yeast Schizosaccharomyces pombe SF3B1 ortholog abolishes its function. (A) Growth of prp10-1 mutant cells expressing wild-type or mutated versions of SpSAP155. prp10-1 mutant cells were transformed with the following yeast expression plasmids: pREP41, pREP41-HA-SAP155, pREP41-HA-SAP155K700E, pREP41-HA-SAP155ins, or pREP41-HA-SAP155K700Eins. Ten-fold serial dilutions of yeast transformants were deposited on EMM 2% glucose media and cultured at 25 °C or 37 °C. (B) RT-PCR splicing analysis of TFIID, NDA3, and RPL7 transcripts in the prp10-1 strain expressing different versions of SpSAP155. Transformants were cultured at 26 °C and subjected to a temperature switch at 37 °C for 2 h. RNA was extracted and analyzed by RT-PCR using specific primers allowing the detection of both spliced and unspliced mRNA. Data information: In (A,B), representative results of three independent experiments are presented. L: ladder.