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Gene/Clone	Species	Stage	Anatomy Item	Experimenter
prox1	xenopus

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Experiment details for prox1

Wu HY et al. (2009) Assay

The role of Xenopus Rx-L in photoreceptor cell determination.

Gene	Clone	Species	Stages	Anatomy
prox1		xenopus	NF stage 35 and 36	lens , retina , eye , retinal inner nuclear layer

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Fig. 4. Effect of Rx-L gain-of-function on the eye development. (A, B) WMISH analysis of embryos Rx-L-GR injected at the 4-cell stage into one of the dorsoanimal blastomeres. (Aa–j) Anterior views of embryos (dorsal up; injected-side to the right) probed with antisense RNA of Rx1 (Aa–e) or Six3 (Af–j). (Aa, f) NF stage 16 embryos not treated with dexamethazone (Dex). (Ab, g) Embryos treated with Dex at NF stage 14, neither Rx1 nor Six3 expression was affected till NF stage 16. (Ac, h) At NF stage 24, Rx-L-GR injected embryos treated with Dex at NF stage 14 show reduced expression domains of Rx1 and Six3. (Ad, i, e, j) Rx-L-GR injected embryos treated with Dex at NF stage 17, here, the domains of Rx1 and Six3 expression were extended at NF stage 19 and 24. (B) Transversal retinal sections of NF stage 36 embryos stained for maker genes of different retinal cell types. (Ba–e) Retinae of Rx-L-GR injected embryo not treated with Dex. (Ba'–e') Retinae of embryos injected with Rx-L-GR mRNA and treated with Dex at NF stage 17/18. Induction of Rx-L-GR function not only led to Rhodopsin expression (Ba,Ba') in folds evaginated from the ONL into the INL, or the GCL (Ba', arrows), but also to the discrete spots of Rhodopsin expression (Ba' arrowhead). Activation of Rx-L-GR reduced the density of the expression of horizontal cell marker, Prox1 (Bb, Bb'), bipolar cell marker, Vsx1 (Bc, Bc'), and amacrine cell marker Pax6 (Bd, Bd'), while the expression of the ganglion cell marker, Brn3.0 (Be, Be') was not significantly changed (Be, Be'). (C) Effects of Rx-L-GR microinjection on apoptosis and cell proliferation. (Ca–c) Transversal sections (dorsal sides up and injected sides to the right) of embryos injected with 50 pg Rx-L-GR (+Dex at NF stage 17) into one of the dorsoanimal blastomeres at the 4-cell stage and subjected to TUNEL assay (Ca) or immunostaining of phosphorylated histone H3 (p-Histone 3, Cb, c) at NF stage 34 and NF stage 24 to detect apoptotic or mitotic cells, respectively. (Ca'–c') Graphs showing the numbers of TUNEL positive cells (Ca') or p-Histone 3 positive cells (C', c') in total or eye area of the non-injected side (green bars), and Rx-L-GR injected side (blue bars). The average of TUNEL or p-Histone-3 positive cell numbers on per section was determined in each embryo. In the TUNEL assay, for non-injected side, n = 5 embryos; for Rx-L-GR injected side, n = 2 embryos. For p-Histone-3 detection, non-injected side, n = 2 embryos; for Rx-L-GR injected side, n = 2 embryos at each stage. Values are given as means ± S.E.M. p = 0.28, , p = 0.07, compared with the non-injected side (Student's t-test). Quantification of the counted cells and sections is shown next to the graphs respectively.

Gene	Clone	Species	Stages	Anatomy
prox1		xenopus	NF stage 37 and 38	lens , retina , eye , retinal inner nuclear layer

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Fig. 3. Rx-L function is needed after eye field induction. (A, B) WMISH analysis of staged embryos injected with MoRx-L at 4-cell stage into one of the dorsoanimal blastomeres. (Aa–Af) Frontal views, injected side to the right. At NF stage 15/16, the expressions of Pax6 (Aa), Rx1 (Ab) and Six3 (Ac) are not significantly affected due to suppression of Rx-L function. (Ad–Af) At tailbud stage (NF stage 24), the expressions of Pax6 (Ad), Rx1 (Ae) and Six3 (Af) are all reduced in the injected sides. (Ba–f') Transversal gelatin/albumin sections of injected embryos probed with the markers of different retinal cell types, with dorsal side upward. The staining of each probe in the retina of injected side (Ba'–f') should be compared with that in the corresponding control side of the same embryo (Ba–f) respectively. The red dashes in Ba' mark the presumptive RPE. Suppression of Rx-L function led to a dramatically reduced expression of the photoreceptor markers, Rhodopsin (Ba, Ba') and Arrestin-C (Bb, Bb'), and a broadened expression of the bipolar cell maker, Vsx1 (Bd, Bd') and the amacrine/ganglion cell marker, Pax6 (Be, Be'). The expression levels Prox1 (Bc, Bc') and Brn3.0 (Bf, Bf') remain almost unchanged upon MoRx-L injection. (C) Effects of MoRx-L microinjection on apoptosis and cell proliferation. (Ca, b, d, e) Transversal sections (dorsal sides up and injected sides to the right) of embryos injected with 2.5pmol of MoRx-L (Ca, d) or Contr-Mo (Cb, e) subjected to TUNEL assay (Ca, b) or immunostaining of phosphorylated histone H3 (p-Histone 3) (Cd, e) at NF stage 31/32 to detect apoptotic or proliferating cells, respectively. (Cc, f) Graphs showing the numbers of TUNEL positive cells (Cc) or p-Histone 3 positive cells (Cf) in total or eye area of the non-injected side (green bars), Contr-Mo injected side (green bars), and MoRx-L (orange bars). The average of TUNEL or p-Histone-3 positive cell numbers on per section was determined in each embryo. In the TUNEL assay, for non-injected, n = 5 embryos; for Contr-Mo, n = 2 embryos; for MoRx-L, n = 3 embyros. In the p-Histone 3 detection, for non-injected, n = 5 embryos; for Contr-Mo, n = 2 embryos; for MoRx-L, n = 3 embyros. Values are given as means ± S.E.M. p = 0.28, , p < 0.05, compared with the non-injected side (Student's t-test). Quantification of the counted cells and sections is shown next to the graphs respectively.