JCS03420 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. The JNK pathway is involved in crystal cell activation. (A) A group of bsk mutant hemocytes (act-Gal4>UAS-bsk-RNAi) 1 hour after bleeding with one crystal cell (indicated by the lower arrows) where crystals are still visible and a second crystal cell which has not ruptured but the crystals are partially dissolved (upper arrow). (B,C) Clot from bsk mutants in phase contrast (B) and normal light (C) showing the lack of melanization. (D,E) Crystal cells visualized using heating (as in Fig. 1) in a bsk RNAi mutant larva (D) and in a control larva (E: UAS-bsk/TM6B, Tb¹). (F,G) The JNK pathway is activated in crystal cells. Larvae with a LacZ reporter under the control of the puckered control region were analyzed using phase contrast (F) and normal light (G). Bars, 10 μm.

• Supplemental Figure 2 -

  Fig. S2. Activation of crystal cells is defective in RhoA mutants. (A) Phase contrast and (B) bright field micrograph of a hemolymph clot from larvae expressing a dominant active form of RhoA (RhoA-DA) (Billuart et al., 2001) in hemocytes at 29°C. Expression of RhoA-DA in hemocytes was induced using the hemocyte specific hemese-GAL4 driver (Zettervall et al., 2004). Note the almost complete lack of melanization and the more rounded appearance of plasmatocytes in the clot. (C-E) A crystal cell from RhoA-DA mutant larva at 25°C analyzed directly (C) and 60 minutes (D, phase contrast and E, bright field) after bleeding. Crystal cells fail to release the PO-containing crystal (arrow). (F,G) Crystal cells in mutant larvae at 25°C show less melanization. Larvae were heat-treated (like in Fig. 1) leading to PO activation in crystal cells and strong melanization in the control (F) and much weaker activity in mutant larvae. Bars, 10 μm.

• Movie 1 -

  Movie 1. Crystal cell rupture. Hemocytes were recorded between 1 and 11 minutes after bleeding. The rupture of a crystal cell and dissolution of the phenoloxidase-containing crystal are visible (see also Fig. 1) as well as two plasmatocytes.