Fig 3. Quantification of
lin-22 expression in the seam. (A) Transgenic animals carrying transcriptionalreporters consisting of various fragments of upstream of
lin-22 sequences fused to GFP. From top to bottom:full
lin-22 endogenous promoter, distal
lin-22 promoter region that is deleted in
lin-22(
icb38), proximal
lin-22promoter present in
lin-22(
icb38), distal
lin-22 promoter with deleted CR1, and CR1 only driving expression ofGFP. White arrows indicate expression in the seam cells; white arrowheads expression in the hypodermis andgreen arrowheads expression in intestinal cells. (B) Quantification of expression pattern for eachtranscriptional reporter (n=35 animals). (C) Representative smFISH images showing
lin-22 expression(black spots correspond to mRNAs and seam cells are labelled in green due to scm::GFP expression) in wildtype V cells after the symmetric L2 division (top), the L2 asymmetric division (middle), and late after the L2asymmetric division (bottom). (D) Quantification of
lin-22 spots per seam cell in wild-type and
lin-22(
icb38)animals at the late L1 stage (n 10 cells per genotype). (E) Quantification of
lin-22 spots in wild-type,
lin-22(
ot267),
lin-22(
ot269), and
lin-22(
icb49) mutants in pools of H cells and V cells at the late L1 stage (n 41).(F-G) Comparison of number of
lin-22 spots between wild-type and the
elt-1(
ku491) mutant (F) or the
egl-18(
ok290) mutant, (G) both at the late L1 stage in pools of H and V cells (n 49). Black stars show statisticallysignificant changes in the mean with a t test or one-way ANOVA as follows: * P < 0.05, ** P < 0.01,*** P < 0.001, **** P < 0.0001. Reference samples for comparisons in E, F, G are the control samplesdepicted in black. Scale bars in A and C are 100 μm and 10 μm, respectively. Error bars show mean +- SEM(D, F, G) or mean ± SD (E). Numerical data used for Fig 3B, D, E, F, G can be found in S2 Data. CR1,conserved region 1; GFP, green fluorescent protein; L1, first larval stage; L2, second larval stage; smFISH,single molecule fluorescent in situ hybridization.