Laminin subunits and their role in C. elegans development

DEV00481 Supplemental Data

Files in this Data Supplement:

• Figure S1 -

  Fig. S1. The localization of laminin aA and aB is independent of each other. (A) The lam-3(n2561) mutants lack laminin aA. Staining with Laminin aB antibodies indicates that Laminin aB is localized to the pharyngeal (p), intestinal (i), and gonadal (g) basement membranes as in wild type.  (B) The epi-1(rh199) mutants lack laminin aB. Staining with laminin aA antibodies indicates that laminin aA is localized to the pharyngeal (p) basement membrane as in wild type.

• Figure S2 -

  Fig. S2. The laminin a subunit antibodies are specific. (A) A lam-3(n2561)/+ embryo (left) and a lam-3(n2561)/lam-3(n2561) embryo (right) stained with MH4 antibodies. (B) The same embryos stained with anti-laminin aA. No specific staining is detected in the lam-3(n2561)/lam-3(n2561) embryo (right). (C) A epi-1(rh199)/+ embryo (left) and a epi-1(rh199)/ epi-1(rh199) embryo (right) stained with anti-UNC-54 antibodies. (D) The same embryos stained with anti-laminin aB antibodies. No specific staining is detected in the epi-1(rh199)/ epi-1(rh199) embryo (right).

• Figure S3 -

  Fig. S3. The initial localization of Laminin aA and aB is normal in emb-9 collagen IV mutant embryos. (A) emb-9(gm23) were stained during early elongation using Laminin aA antibodies. The embryo on the left is at a terminal stage of development. As in wild-type embryos, laminin aA is associated with the developing pharyngeal (arrows) and intestinal basement membranes (arrowheads). (B) emp-9(gm23) embryos stained during elongation using aB antibodies.  As in wild-type embryos, laminin aB is associated with muscle basement membranes (arrows).

• Figure S4 -

  Fig. S4. Morphology of C. elegans basement membranes. (A) Body wall muscle (m) and epidermis (e) have thin sheets of basement membranes surrounding them. The epidermal basement membrane has a unique ‘lollypops’ appearance (arrowhead) with a row of dark dots sticking out from one side of the basement membrane. The outward-facing body wall muscle basement membrane (white arrow) has a thicker appearance than the inward-facing basement membrane (black arrow). (B) The pharyngeal marginal (ma) and muscle (m) cells are radially organized about the lumen (asterisk). The basement membrane surrounding the pharynx has a very thick appearance (arrows). (C) The specialized distal tip cell (dtc) embraces the distal tip of the gonad and extends long thin processes to cover part of the germ cells (g). The gonad and intestine (i) are covered by separate basement membranes. (D) The distal tip cell is covered by a thick basement membrane (arrow), whereas a thinner basement membrane (arrowhead) surrounds the germ cells. No basement membrane lies between these cells. Scale bars: 5 mm in A-C; 1 mm in D.

• Figure S5 -

  Fig. S5. The asymmetry of basement membranes. (A) Gonad sheath (s) covering a developing germ cell (g), where a thin basement membrane (arrow) covers the sheath where it faces the pseudocoelom, but no basement membrane separates the sheath from the germ cell (arrowhead). (B) Gonad sheath covering a more developed oocyte (o) now shows traces of a thick basement membrane (arrowhead) being deposited between sheath and oocyte. A thin basement membrane (arrow) covers the sheath where it faces the pseudocoelom. (C) Between two mature unfertilized oocytes, thick basement membrane (arrowheads) separates each oocyte (o). (D) In mature male tail, thick basement membrane (arrowheads) covers cloacal muscles (c) where they face the cuticle of the gubernaculum (gu), and invaginates between muscles near this contact region, while non contact regions of the cloacal and spicule muscles (sp) are covered by thin basement membranes (arrows). (E) Bodywall muscles (m) have thick basement membrane (arrowheads) separating them from epidermis (e), but have thin basement membranes (arrows) where they face the pseudocoelom or each other. (F) Pharynx (ph) is covered by a thick basement membrane (arrowheads) facing the pseudocoelomic space, while the overlying bodywall tissue (epidermis) has a thin basement membrane (arrows).