Figure 4 Rescue of cell migration defects in
sdn-1 mutants. (A-C) Fluorescent images of animals expressing the translational reporter Psdn-1::
sdn-1::GFP. (A) Expression of SDN-1::GFP is seen in the ALM and AVM mechanosensory neurons. Expression was also observed in the motor neurons (mn),motor neuron commissures (mnc), dorsal nerve cord (dnc), and ventral nerve cord (vnc). (B and C) SDN-1::GFP expression is seen in the HSN neurons.Expression was also detected in the motor neuron commissures (mnc) and in the hypodermis around the seam cells (arrowheads). Bars indicate 20 um.(D) Distribution of ALM cell positions in wild-type (WT) control (black dashed line),
sdn-1(
zh20) mutant (gray dashed line), and
sdn-1(
zh20) mutant animals with a representative rescuing transgene (red dashed line). The X axis shows the ratio of the distance from the cell body to the anus divided bythe length of the animal from the nose to the anus to indicate cell position along the anterior-posterior axis of the animal. All animals contained thezdIs5 I [Pmec-4::GFP] transgene to visualize ALM neurons, a
pha-1(
e2123) mutant allele, and transgene containing
pha-1 rescuing DNA plus additionaltransgenes as indicated. Statistical comparisons were calculated using the Kolmogorov-Smirnov test and are indicated in black (for comparison againstWT) and gray (for comparison against
sdn-1(
zh20) mutant animals). Asterisks denote statistical significance: *** P , 0.0005; ns, not significant. N $22 for all lines analyzed. Data for two additional transgenic lines is shown in File S4. (E) Distribution of HSN cell positions (shown as the percentage ofHSNs in bins as indicated) in
sdn-1(
zh20) mutant animals containing a representative rescuing transgene (+) or their nontransgenic siblings (2). For acomparable WT and mutant distribution, see Figure 2C. Statistical comparisons were calculated using Fisher's exact test and are indicated as: * P , 0.05.N $ 50. Data for seven additional transgenic lines is shown in File S2. (F) The percentage of animals with defects in coelomocyte migration in threeindependent transgenic lines carrying a rescuing transgene (+) or their nontransgenic siblings (2) (gray bars) are shown with error bars representing theSE of proportion. The defect in
sdn-1(
zh20) mutant animals (black bar) is shown for comparison only and identical to data shown in Figure 2D. Asterisksdenote statistical significance: ** P , 0.005, ns, not significant. N $ 86. (G and H) Distributions of ALM (G) and AVM (H) cell positions in
sdn-1(
zh20);
pha-1 mutant animals. Schematics show the cell migration within the animal as well as the SDN-1 primary protein structure with domains indicated (tm:transmembrane). All animals carry transgenes containing the
pha-1 rescuing DNA (pBX) alone (labeled "
sdn-1(
zh20)" in gray), or a construct with theunc-119 promoter driving a wild-type
sdn-1 cDNA (labeled "WT control" in black) or mutant versions of the
sdn-1 cDNA as indicated (labeled e.g.,"S71A," in red, blue, or green for independent transgenic lines). The number of rescuing lines (defined as statistically different from "
sdn-1(
zh20)"mutant but not "WT control") out of the total number of lines is indicated below each point mutant. The X axis shows the ratio of the distance from thecell body to the anus divided by the length of the animal from the nose to the anus to indicate cell position along the anterior-posterior axis of theanimal. All animals contained the zdIs5 I [Pmec-4::GFP] transgene to visualize ALM and AVM neurons. Statistical comparisons between individual linesand "WT control" or "
sdn-1(
zh20)" mutant data (shown in the respective top panels) were calculated using the Kolmogorov-Smirnov test. Asterisksdenote statistical significance: * P , 0.05, ** P , 0.005, *** P , 0.0005; ns, not significant, and are color coded in black (for comparisons with "WTcontrol") and gray (for comparisons with "
sdn-1(
zh20)"). N $ 22 for all lines analyzed. Data for all transgenic lines is shown in File S4.