Supplementary figures

Supplementary figures

Files in this Data Supplement:

• Supplementary figure 1 - ATM phospho substrate is detected at elevated levels in TSC at mitosis. TSC were cultured, fixed, and developed by immunocytochemical means for ATM phospho substrate. (A, B, C) TSC stained with ATM phospho substrate stain, Hoechst stain, and after merging[q;1; is ‘after merging’ here and in subsequent legends OK?] respectively. TSC in anaphase with elevated expression localized in the apparent mitotic spindle are shown by an arrow in A and arrowheads in B and C. (D) No-antibody control; (E) the respective Hoechst-stained nuclei.
• Supplementary figure 2 - MycB phospho is detected at elevated levels in TSC at mitosis. TSC were cultured, fixed, and developed by immunocytochemical means for MycB phospho. (A, B, C) TSC stained with MycB phospho stain, Hoechst stain, and after merging respectively. TSC in apparent metaphase with elevated expression localized in the apparent mitotic spindle are shown by an arrow in A and arrowheads in B and C. (D) No-antibody control; (E) the respective Hoechst-stained nuclei.
• Supplementary figure 3 - p53 phospho is detected at elevated levels in TSC at mitosis. TSC were cultured, fixed, and developed by immunocytochemical means for p53 phospho. (A, B, C) TSC stained with p53 phospho stain, Hoechst stain, and after merging respectively. TSC in apparent anaphase with elevated expression localized in the apparent mitotic spindle are shown by an arrow in A and arrowheads in B and C. (D) No-antibody control; (E) the respective Hoechst-stained nuclei.
• Supplementary figure 4 - p57 all forms is detected at elevated levels in TSC at interphase. TSC were cultured, fixed, and developed by immunocytochemical means for p57 all forms. (A, B, C) TSC stained with p57 all forms (SC8298) stain, Hoechst stain, and after merging respectively. TSC in apparent anaphase with elevated expression localized in the apparent mitotic spindle are shown by an arrow in A and arrowheads in B and C. (D) No-antibody control; (E) the respective Hoechst-stained nuclei. F, G, H, I, J are in the same sequence as A, B, C, D, E respectively but with a second p57 antibody, MS-1062.
• Supplementary figure 5 - Ets2 all forms is detected at elevated levels in TSC at mitosis. TSC were cultured, fixed, and developed by immunocytochemical means for Ets2 all forms. (A, B, C) TSC stained with Ets2 all forms stain, Hoechst stain, and after merging respectively. TSC in apparent metaphase with elevated expression localized in the apparent mitotic spindle are shown by an arrow in A and arrowheads in B and C. (D) No-antibody control; (E) the respective Hoechst-stained nuclei.
• Supplementary figure 6 - Threonine (Thr) phospho is detected at elevated levels in TSC at mitosis. TSC were cultured, fixed, and developed by immunocytochemical means for Thr phospho. (A, B, C) TSC stained with Thr phospho stain, Hoechst stain, and after merging respectively. TSC in apparent metaphase with elevated expression localized in the apparent mitotic spindle are shown by an arrow in A and arrowheads in B and C. (D, E, F) E3.5 embryos stained with Thr phospho stain, Hoechst stain, and after merging respectively. (G, H) No-antibody control for TSC and the corresponding Hoechst stain; (I, J) the same controls for embryos.[q;2; explain arrow and arrowheads in D-F]
• Supplementary figure 7 - MEK1, 2 phospho and p27 phospho proteins are detected at elevated levels in TSC and E3.5 embryos at mitosis. MEK1, 2 phospho proteins are detected at elevated levels in TSC and E3.5 embryos at apparent prophase and anaphase mitosis. TSC were cultured, fixed, and developed by immunocytochemical means for MEK1, 2 phospho. (A, B, C) TSC stained with MEK1, 2 phospho stain, Hoechst stain, and after merging respectively. TSC in apparent metaphase with elevated expression localized in the apparent mitotic spindle are shown by the bottom arrow in A while the top arrow shows a putative prophase cell with passenger MEK1, 2 phospho spots on the DNA. Corresponding arrowheads show nuclear position in B and C. (D, E, F) E3.5 embryos stained with MEK1, 2 phospho stain, Hoechst stain, and after merging respectively. (D) Arrow shows putative M phase cell with high MEK1, 2 phospho and arrowheads show nuclei (E, F). (G, H, I) Karyomax-treated TSC, and uniform nuclear spotting of MEK1, 2 phospho, the corresponding Hoechst stain, and after merging respectively. (J, K, L) The same p27 phospho stain, Hoechst stain, and after merging respectively. (M, N, O, P) No-antibody controls and respective Hoechst-stained pair for the cells and embryos.[q;3; explain arrows and arrowheads in G-L]
• Supplementary figure 8 - RSK1 phospho is detected at elevated levels in TSC at mitosis. TSC were cultured, fixed, and developed by immunocytochemical means for RSK1 phospho. (A, B, C) TSC stained with RSK1 phospho stain, Hoechst stain, and after merging respectively. Three TSC in apparent metaphase with elevated expression localized in the apparent mitotic spindle are shown by an arrow in A and arrowheads in B and C. (D, E, F) E3.5 embryos stained with RSK1 phospho stain, Hoechst stain, and after merging respectively. An arrow shows a cell in apparent M phase (D) and arrowheads show nuclear position (E, F). (G, H) No-antibody control for TSC and corresponding Hoechst stain; (I, J) the same controls for embryos.
• Supplementary figure 9 - RSK3 phospho is detected at elevated levels in TSC at mitosis. TSC were cultured, fixed, and developed by immunocytochemical means for RSK3 phospho. (A, B, C) TSC stained with RSK3 phospho stain, Hoechst stain, and after merging respectively. TSC in apparent telophase with elevated expression localized in the apparent mitotic spindle are shown by an arrow in A and arrowheads in B and C. (D, E, F) E3.5 embryos stained with RSK3 phospho stain, Hoechst stain, and after merging respectively. An arrow shows a cell in apparent M phase (D) and arrowheads show nuclear position (E, F). (G, H) No-antibody control for TSC and corresponding Hoechst stain; (I, J) the same controls for embryos.
• Supplementary figure 10 - p38MAPK phospho is detected at elevated levels in TSC at mitosis. TSC were cultured, fixed, and developed by immunocytochemical means for p38MAPK phospho. (A, B, C) TSC stained with p38MAPK phospho stain, Hoechst stain, and after merging respectively. Two TSC in apparent metaphase with elevated expression localized in the apparent mitotic spindle poles are shown by arrows in A and arrowheads in B and C. (D, E, F) E3.5 embryos stained with p38MAPK phospho stain, Hoechst stain, and after merging respectively. An arrow shows a cell in apparent M phase (D) and arrowheads show nuclear position (E, F). (G, H) No-antibody control for TSC and corresponding Hoechst stain; (I, J) the same controls for embryos.
• Supplementary figure 11 - MycC phospho is detected at elevated levels in TSC at mitosis. TSC were cultured, fixed, and developed by immunocytochemical means for MycC phospho. (A, B, C) TSC stained with MycC phospho stain, Hoechst stain, and after merging respectively. Two TSC in apparent metaphase with elevated expression localized in the apparent mitotic spindle poles are shown by arrows in A and arrowheads in B and C. (D, E, F) E3.5 embryos stained with MycC phospho stain, Hoechst stain, and after merging respectively. An arrow shows a cell in apparent M phase (D) and arrowheads show nuclear position (E, F). (G, H) No-antibody control for TSC and the corresponding Hoechst stain; (I, J) the same controls for embryos.
• Supplementary figure 12 - Cdx2 phospho is detected at elevated levels in HTR at mitosis. TSC were cultured, fixed, and developed by immunocytochemical means for Cdx2 phospho. (A, B, C) TSC stained with Cdx2 phospho stain, Hoechst stain, and after merging respectively. Two TSC in apparent metaphase with elevated expression localized in the apparent mitotic spindle poles are shown by an arrow in A and arrowheads in B and C. (D, E, F) E3.5 embryos stained with Cdx2 phospho stain, Hoechst stain, and after merging respectively. Arrows show cells in apparent M phase with apparent passenger cCdx2 spots (D) and arrowheads show nuclear position (E, F). (G, H) No-antibody control for TSC and the corresponding Hoechst stain; (I, J) the same controls for embryos.