Fig 5. Gene expression changes associated with loss and gain of daughter cell fate symmetry in
lin22 mutants. (A-B) Representative smFISH images and quantification of wild-type and
lin-22(
icb38) animalsat the L2 symmetric division stage using an
elt-1 probe. (A) Comparable amounts of
elt-1 spots are detected inwild-type V cell daughters and more spots in the posterior H2 daughter. In
lin-22(
icb38) animals, more spotsare detected in the posterior V daughters than the anterior (marked by arrowheads) and even numbers in
the2 H2.p daughters (arrow points to anterior H2.p daughter cell). (B) Quantification of
elt-1 expression in H2.pdaughters (n > 9) and pools of V.p daughter cells (n > 41) of wild-type and
lin-22(
icb38) animals at the L2symmetric division stage. (C)
mab-5 expression expands to posterior daughters of V1-V4 cells (arrowheads)in
lin-22(
icb38) animals, reminiscent of the expression in the posterior V5 cell in wild-type (arrow). (D-F) egl-18smFISH images and quantification of wild-type and
lin-22(
icb38) animals. (D) Quantification of
egl-18 smFISHspots in the H2.p cell daughters at the L2 stage (n 21). (E) Images depicting
egl-18 expression at the L3stage. Note expression in anterior daughter cells in
lin-22(
icb38) animals (arrowheads). (F) Quantification ofegl-18 smFISH spots in V1-V4 cells (n 68) of wild-type and
lin-22(
icb38) animals at the L2 asymmetricdivision stage. (G) Representative
eff-1 smFISH images of wild-type and
lin-22(
icb38) animals at the L3asymmetric division stage. Note absence of signal in the most anterior of the 4 daughter cells in
lin-22(
icb38)animals (arrowheads). H2 consists of 4 cells that have arisen due to symmetric division at the L2 stage. (H)Quantification of seam cell number in wild-type (n = 39),
lin-22(
icb38) (n = 29),
eff-1(
hy21) (n = 31), and
lin-22(
icb38);
eff-1(
hy21) (n = 35) animals. Note that the
eff-1(
hy21) does not show a significant difference in seamcell numbers compared to wild-type, but the double mutant does in comparison to both parental strains. Blackstars show statistically significant changes in the mean with a t test or one-way ANOVA /Dunnett's test. Scalebars in A, C, E, and G are 10 um; black spots correspond to mRNAs and green labels the seam cell nuclei.Error bars in B, D, F, H show mean +- SD. Numerical data used for Fig 5B, D, F, H can be found in S2 Data.GFP, green fluorescent protein; L2, second larval stage; L2sym, symmetric first division at the L2 stage; L3,third larval stage; SCM, seam cell marker; smFISH, single molecule fluorescent in situ hybridization.