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. 2017 Oct 1;28(20):2600–2608. doi: 10.1091/mbc.E17-04-0263

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© 2017 Anthony et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 3: — IMPDH filament assembly does not alter guanine nucleotide biosynthesis. (A) HEK293 cells transfected with the indicated constructs were visualized with antibodies against IMPDH2-myc, total IMPDH, or a DNA stain (DRAQ5). S275L assembles into filaments while Y12A remains diffusely cytoplasmic. One sample was treated with the IMPDH inhibitor ribavirin. (B) IMPDH immunoblot showing endogenous protein expression (lower band) and myc-tagged IMPDH2 (upper band). (C) Incorporation of [¹³C₂ ¹⁵N]glycine into AMP and GMP. Bars denote mean and standard error for three biological replicates conducted on different days. No statistical differences (two-sided Student’s t test; p > 0.05) were found between samples for incorporation of [¹³C₂, ¹⁵N]glycine into AMP pools. (D) Immunofluorescence localization of myc-IMPDH2 (red) and DAPI (blue). (E) Representative IMPDH or GAPDH immunoblot. (F) [¹³C₂ ¹⁵N]glycine incorporation into AMP and GMP as in C. No statistically significant differences were found (two-sided Student’s t test; p > 0.05).