Related to Fig 3.

<p>(A) Maximum intensity projection of the distal germ line of a late L4 larvae showing DAF-2::AID::mNG (green) and germ cell membranes (magenta; tagRFP::PH). DAF-2::AID::mNG is shown in inverted grey scale below. DAF-2::AID::mNG in the distal germ line is predominantly found in cytoplasmic puncta (arrow heads) in GSPCs and in the rachis (the shared inner core of cytoplasm to which all germ cells are connected via cytoplasmic bridges). Scale bar = 10 μm. (B) The mean DAF-2::AID::mNG fluorescence intensity per distal germ line, normalized to control animals (no TIR1). Dots represent the mean value per each gonad arm and one gonad arm was assessed per animal. Bar plots show the mean with error bars showing the standard deviation. DAF-2::AID::mNG levels are lower in <i>sun-1p</i>::<i>TIR1</i> animals without auxin treatment. DAF-2::AID::mNG levels in <i>eft-3p</i>::<i>TIR1</i> germ lines without auxin treatment are not different from control, but are higher compared to <i>sun-1p</i>::<i>TIR1</i> germ lines without auxin. Auxin treatment does not deplete DAF-2::AID::mNG in <i>lim-7p</i>::<i>TIR1</i> or <i>ges-1p</i>::<i>TIR1</i> germ lines, with slightly elevated levels in <i>ges-1p</i>::<i>TIR1</i> germ lines following auxin treatment. (C) Maximum intensity projections showing DAF-2::AID::mNG in the germ line (outlined in orange) and developing vulva (blue arrow head) in control animals (no TIR1) and in animals carrying <i>sun-1p</i>::<i>TIR1</i> and <i>eft-3p</i>::<i>TIR1</i> without auxin treatment, and in <i>lim-7p</i>::<i>TIR1</i> and <i>ges-1p</i>::<i>TIR1</i> with auxin treatment. The vulva is shown as an example of somatic depletion in <i>eft-3p</i>::<i>TIR1</i>. Scale bar = 10 μm. (D) GSPC mitosis is delayed in animals after <i>daf-2</i> was depleted by RNAi throughout the whole worm but not when <i>daf-2</i> was depleted in the germ line alone. Small dots represent individual GSPCs, larger dots represent the mean value per gonad arm/PZ. Boxplots show the median, interquartile range and most extreme values not considered statistical outliers. (E) Lower magnification, whole worm images showing partial DAF-2::AID::mNG depletion in <i>eft-3p</i>::<i>TIR1</i> animals without auxin treatment. Orange arrow heads indicate prominent sites of somatic DAF-2::AID::mNG expression (head neurons, vulva, intestine) in control animals (no TIR1; left) and the same anatomical locations in animals bearing <i>eft-3p</i>::<i>TIR1</i> showing reduced expression. Scale bar = 50 μm. For all plots, n.s. = <i>p</i> > 0.05, * = <i>p</i> < 0.05, *** = <i>p</i> < 0.001. Summary statistics and statistical tests used for all figure panels are given in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1011351#pgen.1011351.s010" target="_blank">S2 Data</a>.</p> <p>(PDF)</p>