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Quantification of immature cortical thymocytes . Stage 60 X. laevis metamorphic tadpoles were exposed to 1 μg/L of the mixture for 21 days. At days 7, 14 and 21 post-metamorphosis, froglet thymocytes were evaluated for CD8, CTX (cortical thymocyte-specific antigen of Xenopus ) and <t>EdU</t> <t>(5-ethynyl-2′-deoxyuridine)</t> fluorescence via flow cytometry. Representative gating for the frequency of CD8 + and CTX+ ( A ) and proliferative EdU + CD8+/CTX + thymocytes ( B ) are shown. The CD8+/CTX + thymocytes were then quantified for total cell number ( C ) and total number of proliferative EdU + CD8+/CTX + thymocytes ( D ). N = 4 independent samples of 3 pooled tadpoles, * denotes statistical significance using 2-way ANOVA and Tukey’s post-test.
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Quantification of immature cortical thymocytes . Stage 60 X. laevis metamorphic tadpoles were exposed to 1 μg/L of the mixture for 21 days. At days 7, 14 and 21 post-metamorphosis, froglet thymocytes were evaluated for CD8, CTX (cortical thymocyte-specific antigen of Xenopus ) and <t>EdU</t> <t>(5-ethynyl-2′-deoxyuridine)</t> fluorescence via flow cytometry. Representative gating for the frequency of CD8 + and CTX+ ( A ) and proliferative EdU + CD8+/CTX + thymocytes ( B ) are shown. The CD8+/CTX + thymocytes were then quantified for total cell number ( C ) and total number of proliferative EdU + CD8+/CTX + thymocytes ( D ). N = 4 independent samples of 3 pooled tadpoles, * denotes statistical significance using 2-way ANOVA and Tukey’s post-test.
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Quantification of immature cortical thymocytes . Stage 60 X. laevis metamorphic tadpoles were exposed to 1 μg/L of the mixture for 21 days. At days 7, 14 and 21 post-metamorphosis, froglet thymocytes were evaluated for CD8, CTX (cortical thymocyte-specific antigen of Xenopus ) and <t>EdU</t> <t>(5-ethynyl-2′-deoxyuridine)</t> fluorescence via flow cytometry. Representative gating for the frequency of CD8 + and CTX+ ( A ) and proliferative EdU + CD8+/CTX + thymocytes ( B ) are shown. The CD8+/CTX + thymocytes were then quantified for total cell number ( C ) and total number of proliferative EdU + CD8+/CTX + thymocytes ( D ). N = 4 independent samples of 3 pooled tadpoles, * denotes statistical significance using 2-way ANOVA and Tukey’s post-test.
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Quantification of immature cortical thymocytes . Stage 60 X. laevis metamorphic tadpoles were exposed to 1 μg/L of the mixture for 21 days. At days 7, 14 and 21 post-metamorphosis, froglet thymocytes were evaluated for CD8, CTX (cortical thymocyte-specific antigen of Xenopus ) and <t>EdU</t> <t>(5-ethynyl-2′-deoxyuridine)</t> fluorescence via flow cytometry. Representative gating for the frequency of CD8 + and CTX+ ( A ) and proliferative EdU + CD8+/CTX + thymocytes ( B ) are shown. The CD8+/CTX + thymocytes were then quantified for total cell number ( C ) and total number of proliferative EdU + CD8+/CTX + thymocytes ( D ). N = 4 independent samples of 3 pooled tadpoles, * denotes statistical significance using 2-way ANOVA and Tukey’s post-test.
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Quantification of immature cortical thymocytes . Stage 60 X. laevis metamorphic tadpoles were exposed to 1 μg/L of the mixture for 21 days. At days 7, 14 and 21 post-metamorphosis, froglet thymocytes were evaluated for CD8, CTX (cortical thymocyte-specific antigen of Xenopus ) and <t>EdU</t> <t>(5-ethynyl-2′-deoxyuridine)</t> fluorescence via flow cytometry. Representative gating for the frequency of CD8 + and CTX+ ( A ) and proliferative EdU + CD8+/CTX + thymocytes ( B ) are shown. The CD8+/CTX + thymocytes were then quantified for total cell number ( C ) and total number of proliferative EdU + CD8+/CTX + thymocytes ( D ). N = 4 independent samples of 3 pooled tadpoles, * denotes statistical significance using 2-way ANOVA and Tukey’s post-test.
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Quantification of immature cortical thymocytes . Stage 60 X. laevis metamorphic tadpoles were exposed to 1 μg/L of the mixture for 21 days. At days 7, 14 and 21 post-metamorphosis, froglet thymocytes were evaluated for CD8, CTX (cortical thymocyte-specific antigen of Xenopus ) and <t>EdU</t> <t>(5-ethynyl-2′-deoxyuridine)</t> fluorescence via flow cytometry. Representative gating for the frequency of CD8 + and CTX+ ( A ) and proliferative EdU + CD8+/CTX + thymocytes ( B ) are shown. The CD8+/CTX + thymocytes were then quantified for total cell number ( C ) and total number of proliferative EdU + CD8+/CTX + thymocytes ( D ). N = 4 independent samples of 3 pooled tadpoles, * denotes statistical significance using 2-way ANOVA and Tukey’s post-test.
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Quantification of immature cortical thymocytes . Stage 60 X. laevis metamorphic tadpoles were exposed to 1 μg/L of the mixture for 21 days. At days 7, 14 and 21 post-metamorphosis, froglet thymocytes were evaluated for CD8, CTX (cortical thymocyte-specific antigen of Xenopus ) and <t>EdU</t> <t>(5-ethynyl-2′-deoxyuridine)</t> fluorescence via flow cytometry. Representative gating for the frequency of CD8 + and CTX+ ( A ) and proliferative EdU + CD8+/CTX + thymocytes ( B ) are shown. The CD8+/CTX + thymocytes were then quantified for total cell number ( C ) and total number of proliferative EdU + CD8+/CTX + thymocytes ( D ). N = 4 independent samples of 3 pooled tadpoles, * denotes statistical significance using 2-way ANOVA and Tukey’s post-test.
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Quantification of immature cortical thymocytes . Stage 60 X. laevis metamorphic tadpoles were exposed to 1 μg/L of the mixture for 21 days. At days 7, 14 and 21 post-metamorphosis, froglet thymocytes were evaluated for CD8, CTX (cortical thymocyte-specific antigen of Xenopus ) and EdU (5-ethynyl-2′-deoxyuridine) fluorescence via flow cytometry. Representative gating for the frequency of CD8 + and CTX+ ( A ) and proliferative EdU + CD8+/CTX + thymocytes ( B ) are shown. The CD8+/CTX + thymocytes were then quantified for total cell number ( C ) and total number of proliferative EdU + CD8+/CTX + thymocytes ( D ). N = 4 independent samples of 3 pooled tadpoles, * denotes statistical significance using 2-way ANOVA and Tukey’s post-test.

Journal: Current Research in Toxicology

Article Title: Developmental exposure to thyroid disrupting chemical mixtures alters metamorphosis and post-metamorphic thymocyte differentiation

doi: 10.1016/j.crtox.2022.100094

Figure Lengend Snippet: Quantification of immature cortical thymocytes . Stage 60 X. laevis metamorphic tadpoles were exposed to 1 μg/L of the mixture for 21 days. At days 7, 14 and 21 post-metamorphosis, froglet thymocytes were evaluated for CD8, CTX (cortical thymocyte-specific antigen of Xenopus ) and EdU (5-ethynyl-2′-deoxyuridine) fluorescence via flow cytometry. Representative gating for the frequency of CD8 + and CTX+ ( A ) and proliferative EdU + CD8+/CTX + thymocytes ( B ) are shown. The CD8+/CTX + thymocytes were then quantified for total cell number ( C ) and total number of proliferative EdU + CD8+/CTX + thymocytes ( D ). N = 4 independent samples of 3 pooled tadpoles, * denotes statistical significance using 2-way ANOVA and Tukey’s post-test.

Article Snippet: A subset of 3 developmentally (3 replicates) exposed adult frogs (6 months old) per group was injected with the thymidine analog EdU (5-ethynyl-2′-deoxyuridine; Catalog: A10044, Invitrogen, Waltham MA, USA) via i.p. injection approximately 18 h prior to tissue collection for flow cytometry.

Techniques: Fluorescence, Flow Cytometry