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Effect of Tcl1 expression on ES cell growth.

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posted on 05.08.2013, 03:35 by Tatsushi Miyazaki, Satsuki Miyazaki, Masafumi Ashida, Tomofumi Tanaka, Fumi Tashiro, Jun-ichi Miyazaki

(A) Cell proliferation assay. Cell proliferation between 24 and 48 hours of culture was analyzed by MTT assay. The proliferation rate of Tcl1−/− (KO) was significantly lower than that of wild-type (WT) or Tcl1−/−(CAG-Tcl1) cells. Values are expressed as means ± SD (left panel, n = 12 each; right panel, n = 8 each). Difference from Tcl1−/− (KO) ES cells: ∗∗P<0.01. (B) The percentage of cleaved caspase 3-positive cells was calculated for 9–13 areas selected at random (left panel, n = 9–13, 6312–14211 nuclei per cell line; right panel, n = 10, 3329–4538 nuclei per cell line). The percentage of cleaved caspase 3-positive apoptotic cells in Tcl1−/− (KO) cells was significantly higher than that in wild-type (WT) or Tcl1−/−(CAG-Tcl1) cells. Values are expressed as means ± SD. Difference from Tcl1−/− (KO) ES cells: ∗P<0.05, ∗∗P<0.01. (C) Teratoma formation of wild-type (WT) (n = 9), Tcl1−/− (KO) #4 (n = 6), Tcl1−/−(CAG-EGFP) #5 (n = 3), and Tcl1−/−(CAG-Tcl1) #1 (n = 3) and #3 (n = 5) ES cells. The weight of each teratoma is shown in grams. Horizontal bars indicate the mean of each sample. ∗P<0.03. (D) Histological analysis of the teratomas derived from Tcl1−/− (KO) #4 and #5 and Tcl1−/−(CAG-Tcl1) #1 and #3 ES cells. No histological differences were recognizable between Tcl1−/− (KO) and Tcl1−/−(CAG-Tcl1) ES cells.

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