FIGURE 9.

Expression of MTHFD2L lacking exon 8 in yeast. S. cerevisiae strain MWY4.5 (ser1 ura3 trp1 leu2 his4 ade3–30/65 Δmtd1) was transformed to uracil prototrophy with YEp-rD2L (wild type), YEp-rD2L-x8 (lacking exon 8), or empty vector (YEp24ES). Ura⁺ transformants were streaked onto yeast minimal plates containing serine as a one-carbon donor plus adenine (left) or serine alone (right) and incubated at 30 °C for 4 days. Both plates also contained leucine, tryptophan, and histidine to support the other auxotrophic requirements of MWY4.5. Inset, immunoblot of whole cell lysate from MWY4.5 transformed with the indicated plasmids. Each lane was loaded with 50 μg of protein. The blot was probed with polyclonal antibodies against MTHFD2L (1:1000 dilution).