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Table 2 PAR-2::GFP polarization following osmotic support in EGM (30 min)a

From: The eggshell is required for meiotic fidelity, polar-body extrusion and polarization of the C. elegansembryo

 

150 mM KClb,c

EGM (30 min)d,e

 

no asymmetryf

incorrect/incomplete asymmetryg

correct asymmetryh

no asymmetryf

incorrect/incomplete asymmetryg

correct asymmetryh

control i

0

0

100

0

0

100

gna-2(RNAi)

100

0

0

40

50

10

chs-1(RNAi)

15

85

0

70

20

10

cej-1+B0280.5(coRNAi)

25

63

13

50

50

0

  1. aPAR-2::GFP was examined in single-cell embryos in which 2 discrete Hoechst-staining structures were in close proximity to one another in the cytoplasm. gna-2(RNAi), chs-1(RNAi) and cej-1+B0280.5(coRNAi) fail to extrude the 2nd polar body; therefore, a 3rd Hoechst-staining structure could often be discerned. Numbers are % of embryos displaying phenotype
  2. b gna-2(RNAi), chs-1(RNAi) and cej-1+B0280.5(coRNAi) embryos are osmotically sensitive and burst without osmotic support; therefore, embryos were dissected as quickly as possible in 150 mM KCl (<5 min) and frozen immediately
  3. ccontrol, n = 10; gna-2(RNAi), n = 8; chs-1(RNAi), n = 6; cej-1+B0280.5(coRNAi), n = 24
  4. dembryos were dissected in minimal EGM and incubated at 22°C for 30 min
  5. econtrol, n = 10; gna-2(RNAi), n = 10; chs-1(RNAi), n = 10; cej-1+B0280.5(coRNAi), n = 8
  6. fPAR-2::GFP undetectable or uniform cortical
  7. gPAR-2::GFP lateral or incompletely anterior/posterior. Since loss of function of gna-2, chs-1 or cej-1+B0280.5 prevents polar body extrusion, it was not possible to definitively identify anterior versus posterior
  8. hPAR-2::GFP posterior
  9. iwild type fed OP50