Supplementary information

Supplementary Fig. 2

Detection of Musashi1 expression using an antibody to Musashi1 (clone 14H1).

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Supplementary Fig. 3

Cluster of Musashi1+ cells detected at the initiation site of crypt budding or crypt fission in PTEN mutants.

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Supplementary Fig. 4

Comparison of expression patterns of p-cat-Ser552, Top-Gal, p27^kip1 and cyclin D1.

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Supplementary Fig. 5

Distribution of cells with p-cat-Ser552.

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Supplementary Video 1

PTEN-deficient stem cells initiate crypt fission. Crypts were dissociated from the polyp region of PTEN mutant intestine. Whole-mount staining was performed using two antibodies: anti--cat-S552 (green) to recognize stem cells (normal or abnormal) and anti-cryptdin (red) to recognize Paneth cells. One -cat-S⁵⁵²⁺ cell was located just above the Paneth cells in the proposed stem cell position; the other dividing -cat-S⁵⁵²⁺ cell was found at the tip of the ridge formed between two dividing crypts.

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Supplementary Video 2

PTEN-deficient stem cells initiate crypt budding. Crypts were dissociated from the polyp region of PTEN mutant intestine. Whole-mount staining was performed using two antibodies: anti--cat-^S552 (green) to recognize stem cells (normal or abnormal) and anti-cryptdin (red) to recognize Paneth cells. One -cat-^S552+ cell was located just above the Paneth cells in the proposed stem cell position; a cluster of -cat-^S552+ cells was found at the junction between a budding crypt and the edge of the original crypt.

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Supplementary Methods

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