The Fat Cadherin Acts through the Hippo Tumor-Suppressor Pathway ...
Supplemental Data The Fat Cadherin Acts through the Hippo Tumor-Suppressor Pathway to Regulate Tissue Size Maria Willecke, Fisun Hamaratoglu, Madhuri Kango-Singh, Ryan Udan, Chiao-lin Chen, Chunyao Tao, Xinwei Zhang, and Georg Halder Supplemental References S1. Hamaratoglu, F., Willecke, M., Kango-Singh, M., Nolo, R., Hyun, E., Tao, C., Jafar-Nejad, H., and Halder, G. (2006). The tumoursuppressor genes NF2/Merlin and Expanded act through Hippo signalling to regulate cell proliferation and apoptosis. Nat. Cell Biol. 8, 27–36.
Figure S1. Fat Regulates Cell Proliferation of Uncommitted Precursor Cells Third-instar eye imaginal disc containing ftfd mutant clones that are marked by the absence of GFP expression (green in [A] and [A00 ]). This disc is labeled for BrdU incorporation (red in [A], [A0 ], and [A00 ] and gray in [A000 ]) and stained for the neuronal marker ELAV to detect differentiating photoreceptor cells (blue in [A] and [A0 ]). Wild-type cells arrest in G1 in the morphogenetic furrow (asterisks), and nondifferentiating cells go through one synchronous S phase in the second mitotic wave (arrows). ftfd mutant cell clones show ectopic cell proliferation posterior to the second mitotic wave (arrowheads). These BrdU-incorporating cells are not differentiating photoreceptor cells because they are not expressing ELAV (A0 ). Anterior is to the left.
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Figure S2. Fat Regulates expanded Expression (A and B) Third-instar wing disc homozygous mutant for ft and expressing Ft in the posterior compartment under the control of the hh-Gal4 driver. Disc is stained to detect bGal expression from a lacZ enhancer-trap insertion into the ex gene (red in [A] and [B] and grayscale in [A0 ] and [B0 ]) and to detect the expression of Ft (green in [A] and [B] and grayscale in [A00 ] and [B00 ]). (B) shows a magnification of the pouch region of the disc in (A). Anterior is to the left and dorsal is up for all discs.
Figure S3. Fat Mutant Discs Express Normal Levels of Expanded Western blot probing the amount of Ex expression (upper bands) in third-instar wing imaginal discs from various genotypes as indicated above the blot. The amount of a-Tubulin (lower bands) served as loading controls. Extracts from the following genotypes were loaded: lane 1, ftfd/ft422; lane 2, wt; lane 3, wtsP2/wtsP2; and lane 4, exAP49/ exAP49. Homozygous ft mutant wing discs expressed similar amounts of Ex as the wt control. wts mutants had higher levels of Ex as expected, because wts mutant cells upregulate ex expression [S1]. exAP49 produces a truncated but stable version of Ex that is still recognized by the Ex antibodies [S1]. As for the wts mutant discs, exAP49 mutants also had higher levels of Ex.
Figure S1. Fat Regulates Cell Proliferation of Uncommitted Precursor Cells Third-instar eye imaginal disc containing ftfd mutant clones that are marked by the absence of GFP expression (green in [A] and [A00 ]). This disc is labeled for BrdU incorporation (red in [A], [A0 ], and [A00 ] and gray in [A000 ]) and stained for the neuronal marker ELAV to detect differentiating photoreceptor cells (blue in [A] and [A0 ]). Wild-type cells arrest in G1 in the morphogenetic furrow (asterisks), and nondifferentiating cells go through one synchronous S phase in the second mitotic wave (arrows). ftfd mutant cell clones show ectopic cell proliferation posterior to the second mitotic wave (arrowheads). These BrdU-incorporating cells are not differentiating photoreceptor cells because they are not expressing ELAV (A0 ). Anterior is to the left.
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S2
Figure S2. Fat Regulates expanded Expression (A and B) Third-instar wing disc homozygous mutant for ft and expressing Ft in the posterior compartment under the control of the hh-Gal4 driver. Disc is stained to detect bGal expression from a lacZ enhancer-trap insertion into the ex gene (red in [A] and [B] and grayscale in [A0 ] and [B0 ]) and to detect the expression of Ft (green in [A] and [B] and grayscale in [A00 ] and [B00 ]). (B) shows a magnification of the pouch region of the disc in (A). Anterior is to the left and dorsal is up for all discs.
Figure S3. Fat Mutant Discs Express Normal Levels of Expanded Western blot probing the amount of Ex expression (upper bands) in third-instar wing imaginal discs from various genotypes as indicated above the blot. The amount of a-Tubulin (lower bands) served as loading controls. Extracts from the following genotypes were loaded: lane 1, ftfd/ft422; lane 2, wt; lane 3, wtsP2/wtsP2; and lane 4, exAP49/ exAP49. Homozygous ft mutant wing discs expressed similar amounts of Ex as the wt control. wts mutants had higher levels of Ex as expected, because wts mutant cells upregulate ex expression [S1]. exAP49 produces a truncated but stable version of Ex that is still recognized by the Ex antibodies [S1]. As for the wts mutant discs, exAP49 mutants also had higher levels of Ex.
