Higher ANKRD9 abundance is associated with longer IMPDH2 rods.
a, immunostaining of IMPDH2 in HeLa cells under basal conditions (10% FBS) shows an expected diffuse pattern (left panel). Treatment of these cells with 10 μm ribavirin for 1 h in the absence of recombinant ANKRD9 triggers formation of IMPDH2 rods (right panel). n = 3 for both conditions. Scale bar, 20 μm. b, in the absence of recombinant ANKRD9, nutrient limitation (0.1% FBS for 48 h) causes appearance of short IMPDH2 rods (left panel). Expression of recombinant FLAG-ANKRD9 significantly increases the length of IMPDH2 rods (right panel). n = 3 independent experiments. c, quantification of IMPDH2 rod length in different treatment conditions. The total number of cells analyzed is as follows: n = 150 cells (10% FBS + ribavirin), n = 73 cells (0.1% FBS), and n = 156 cells (0.1% FBS + ANKRD9 expression). Statistical analysis was done using an unpaired t test. ****, p < 0.0001. Error bars represent S.D. Each point is the average rod length of at least 5 cells. d, quantification of the number of rods per cell under nutrient limitation in the absence and presence of recombinant ANKRD9. unpaired t test. ****, p < 0.0001. Error bars represent S.D. Each point is the average rod length of at least 5 cells.