FIG 2 .

The IMD reorganizes along the sidewall of the cell body during PBST starvation. (A) Fluorescence live imaging of the dual IMD marker strain expressing HA-mCherry-GlfT2 (mCh-GlfT2) and Ppm1-mNeonGreen-cMyc (Ppm1-mNG). Exposure of actively growing cells to PBST led to the redistribution of the IMD. Scale bar, 5 µm. (B) Differences in polar fluorescence enrichment between actively growing (log) and 48-h-starved (PBST) cells. Polar enrichment was calculated as the ratio of mean fluorescence intensities between the polar cap and the sidewall region. The black lines indicate the average of 201 cells. (C) Western blotting detection of IMD proteins from sucrose gradient sedimentation demonstrated the biochemical isolation of the IMD in cells starved in PBST for 30 h. IMD markers were HA-mCherry-GlfT2 (anti-HA, 100 kDa), Ppm1-mNeonGreen-cMyc (anti-c-Myc, 59 kDa), and PimB′ (anti-PimB′, 42 kDa). The PM-CW marker was MptA (anti-MptA, 54 kDa). (D) Quantification of alkDADA incorporation, normalized to cell sizes of cells after 30 h of PBST starvation, showing decreased de novo PG synthesis. The black lines indicate the averages of 50 cells. *, P < 0.001.