Figure 4

LI1158 exists as oligomers/monomers in yeast and mammalian cells, respectively. A Expression of recombinant LI1158 is non-toxic in yeast. W303-1A yeast strains carrying the yeast expression vector pRS416-GAL1, expressing either empty vector or LI1158 with an N-terminal 3 × Flag, were grown overnight in repressing medium (2% glucose). Cultures were then normalized to OD₆₀₀ 1.0, and serial tenfold dilutions were grown at 30°C for 2 and 3 days in repressing and inducing medium (2% galactose and 1% raffinose), respectively. W303-1A yeast carrying the yeast expression vector pYES2/NT-RipI was used as the positive control. B LI1158 exists as oligomers in yeast cells. Induction of 3 × Flag-LI1158 expressions at different time points were verified by Western blotting using an anti-Flag antibody. C LI1158 exists as monomers in mammalian cells. Immunoblots of extracts of HEK293T cells transfected with psin-EF1-puro-SFB or pSIN-EF1-puro-SFB-LI1158 for 48 h. Cell lysates were probed with the anti-Flag tag antibody. Asterisks denote some unknown modification bands.