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Supplementary Material

Regulation and function of odd-paired in Tribolium segmentation

Chong Pyo Choe1,2,*, Frank Stellabotte3, and Susan J. Brown4

1 Division of Life Science, Gyeongsang National University, Jinju 52828, Korea

2 Division of Applied Life Science (BK21 Plus Program), Plant Molecular Biology and

Biotechnology Research Center, Gyeongsang National University, Jinju 52828, Korea

3 School of Allied Health, Business, and STEM, Middlesex Community College, Middletown,

CT USA

4 Division of Biology, Kansas State University, Manhattan, KS USA

*Corresponding author: Chong Pyo Choe

Division of Life Science, Gyeongsang National University, Jinju 52828, Korea

Email: cpchoe@gnu.ac.kr, Phone: +82 (055) 772-1367, Fax: +82 (055) 722-1329

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Table S1. Experimental Numbers

Figure Experimental Numbers 2a Expression of Tc-opa in wild-type embryos at the early blastoderm stages (one Tc-opa stripe).

n = 12

2b Expression of Tc-opa in wild-type embryos at the late blastoderm stages (two Tc-opa stripes). n = 18

2c Expression of Tc-opa in wild-type embryos at the early germband stages (early three Tc-opa stripes). n = 31

2d Expression of Tc-opa in wild-type embryos at the early germband stages (late three Tc-opa stripes). n = 37

2e Expression of Tc-opa in wild-type embryos at the elongating germband stages (late four Tc-opa stripes). n = 29

2f Expression of Tc-opa in wild-type embryos at the elongating germband stages (eight Tc-opa stripes). n = 43

2g Expression of Tc-opa in wild-type embryos at the elongating germband stages (eleven Tc-opa stripes). n = 38

2h Expression of Tc-opa in wild-type embryos at the elongating germband stages (thirteen Tc-opa stripes). n = 41

2i Expression of Tc-opa in wild-type embryos at the fully extended germband stages (early sixteen Tc-opa stripes). n = 27

2j Expression of Tc-opa in wild-type embryos at the fully extended germband stages (late sixteen Tc-opa stripes). n = 24

3a Double staining of Tc-opa and Tc-En in wild-type embryos at the elongating germband stages.

n = 34 3b Double staining of Tc-opa and Tc-En in evepRNAi embryos at the elongating germband stages.

n = 37 3c Double staining of Tc-opa and Tc-En in oddpRNAi embryos at the elongating germband stages.

n = 36 3d Double staining of Tc-opa and Tc-En in opapRNAi embryos at the elongating germband stages.

n = 43

S1a Expression of Tc-opa and Tc-En in wild-type embryos at the early blastoderm stages (one Tc-En stripe). n = 16

S1b Expression of Tc-opa and Tc-En in wild-type embryos at the germ rudiment stages (one Tc-En stripe). n = 19

S1c Expression of Tc-opa and Tc-En in wild-type embryos at the elongating germband stages (six Tc-En stripes). n = 23

S1d Expression of Tc-opa and Tc-Eve in wild-type embryos at the elongating germband stages (six Tc-Eve stripes). n = 41

S1e Expression of Tc-opa and Tc-En in wild-type embryos at the fully extended germband stages (fifteen Tc-En stripes). n = 31

S1f Expression of Tc-opa and Tc-En in wild-type embryos at the fully extended germband stages (sixteen Tc-En stripes). n = 26

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Fig. S1 Double staining of Tc-opa (purple) with Tc-En (punctate, dark brown spots) or Tc-Eve

(punctate, brown spots) in Tribolium undergoing embryonic segmentation. a, b Blastoderm

stages. c-f Elongating germband stages. d1-f1 Digitally amplified posterior region of d-f, respectively. a First Tc-opa is co-expressed with the first Tc-En stripe in the mandibular

segment. b When the germ rudiment forms, new Tc-opa expression (arrowhead) is observed

in a broad region posterior to the first Tc-opa stripe. This broad Tc-opa expression occurs

prior to expression of the second Tc-En stripe. c The second Tc-opa stripe is coincident with

the second Tc-Eve stripe (arrowhead) in the growing germband. d Tc-Eve 4b, Tc-Eve 5, and

Tc-Eve 6 stripes identify five segments included within the broad Tc-opa expression domain

(bar) at the growth zone of an elongating germband. The posterior end of the broad Tc-opa

expression domain (arrowhead in d1) is coincident with the Tc-Eve 6 stripe. e, f When the

germband is fully extended, the broad Tc-opa expression domain becomes narrower (bar in e and arrowhead in e1) and is split into the 15th and 16th Tc-opa stripes (arrowheads in f1).

Anterior is to the left.