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SERPINA1-994 <t>decreases</t> <t>polymeric</t> <t>AAT</t> accumulation in human iPSC-derived ZZ hepatocytes. ( A ) Images of cultured iPSCs after 14 days of treatment with the indicated AIMer. AAT polymer staining (green) and DAPI staining of nuclei (blue) are shown. The scale bars are 100 μm. ( B ) Quantification of polymeric AAT after 5, 10, or 14 days of treatment with increasing concentrations (0, 1.25, 2.5, 5.0, or 10 µM) of the indicated AIMer. Data are presented as mean ± SEM ( n = 6). White-adjusted two-way ANOVA with post-hoc comparisons to untreated allowing heteroscedasticity. ( C ) SERPINA1 RNA editing after 5, 10, or 14 days of treatment. Data are presented as mean ± SEM ( n = 4–6). White-adjusted two-way ANOVA with post-hoc comparisons to untreated allowing for heterscedasticity. ( D ) Quantification of nuclei per field after 5, 10, or 14 days. Data are presented as mean ± SEM ( n = 6). Two-way ANOVA with post-hoc comparisons to untreated. For all panels, P = 0.0 indicates that calculated P- values were below the limit of the program (2.2 × 10 −16 ). ns: nonsignificant.
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SERPINA1-994 <t>decreases</t> <t>polymeric</t> <t>AAT</t> accumulation in human iPSC-derived ZZ hepatocytes. ( A ) Images of cultured iPSCs after 14 days of treatment with the indicated AIMer. AAT polymer staining (green) and DAPI staining of nuclei (blue) are shown. The scale bars are 100 μm. ( B ) Quantification of polymeric AAT after 5, 10, or 14 days of treatment with increasing concentrations (0, 1.25, 2.5, 5.0, or 10 µM) of the indicated AIMer. Data are presented as mean ± SEM ( n = 6). White-adjusted two-way ANOVA with post-hoc comparisons to untreated allowing heteroscedasticity. ( C ) SERPINA1 RNA editing after 5, 10, or 14 days of treatment. Data are presented as mean ± SEM ( n = 4–6). White-adjusted two-way ANOVA with post-hoc comparisons to untreated allowing for heterscedasticity. ( D ) Quantification of nuclei per field after 5, 10, or 14 days. Data are presented as mean ± SEM ( n = 6). Two-way ANOVA with post-hoc comparisons to untreated. For all panels, P = 0.0 indicates that calculated P- values were below the limit of the program (2.2 × 10 −16 ). ns: nonsignificant.
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SERPINA1-994 <t>decreases</t> <t>polymeric</t> <t>AAT</t> accumulation in human iPSC-derived ZZ hepatocytes. ( A ) Images of cultured iPSCs after 14 days of treatment with the indicated AIMer. AAT polymer staining (green) and DAPI staining of nuclei (blue) are shown. The scale bars are 100 μm. ( B ) Quantification of polymeric AAT after 5, 10, or 14 days of treatment with increasing concentrations (0, 1.25, 2.5, 5.0, or 10 µM) of the indicated AIMer. Data are presented as mean ± SEM ( n = 6). White-adjusted two-way ANOVA with post-hoc comparisons to untreated allowing heteroscedasticity. ( C ) SERPINA1 RNA editing after 5, 10, or 14 days of treatment. Data are presented as mean ± SEM ( n = 4–6). White-adjusted two-way ANOVA with post-hoc comparisons to untreated allowing for heterscedasticity. ( D ) Quantification of nuclei per field after 5, 10, or 14 days. Data are presented as mean ± SEM ( n = 6). Two-way ANOVA with post-hoc comparisons to untreated. For all panels, P = 0.0 indicates that calculated P- values were below the limit of the program (2.2 × 10 −16 ). ns: nonsignificant.
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SERPINA1-994 <t>decreases</t> <t>polymeric</t> <t>AAT</t> accumulation in human iPSC-derived ZZ hepatocytes. ( A ) Images of cultured iPSCs after 14 days of treatment with the indicated AIMer. AAT polymer staining (green) and DAPI staining of nuclei (blue) are shown. The scale bars are 100 μm. ( B ) Quantification of polymeric AAT after 5, 10, or 14 days of treatment with increasing concentrations (0, 1.25, 2.5, 5.0, or 10 µM) of the indicated AIMer. Data are presented as mean ± SEM ( n = 6). White-adjusted two-way ANOVA with post-hoc comparisons to untreated allowing heteroscedasticity. ( C ) SERPINA1 RNA editing after 5, 10, or 14 days of treatment. Data are presented as mean ± SEM ( n = 4–6). White-adjusted two-way ANOVA with post-hoc comparisons to untreated allowing for heterscedasticity. ( D ) Quantification of nuclei per field after 5, 10, or 14 days. Data are presented as mean ± SEM ( n = 6). Two-way ANOVA with post-hoc comparisons to untreated. For all panels, P = 0.0 indicates that calculated P- values were below the limit of the program (2.2 × 10 −16 ). ns: nonsignificant.
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SERPINA1-994 decreases polymeric AAT accumulation in human iPSC-derived ZZ hepatocytes. ( A ) Images of cultured iPSCs after 14 days of treatment with the indicated AIMer. AAT polymer staining (green) and DAPI staining of nuclei (blue) are shown. The scale bars are 100 μm. ( B ) Quantification of polymeric AAT after 5, 10, or 14 days of treatment with increasing concentrations (0, 1.25, 2.5, 5.0, or 10 µM) of the indicated AIMer. Data are presented as mean ± SEM ( n = 6). White-adjusted two-way ANOVA with post-hoc comparisons to untreated allowing heteroscedasticity. ( C ) SERPINA1 RNA editing after 5, 10, or 14 days of treatment. Data are presented as mean ± SEM ( n = 4–6). White-adjusted two-way ANOVA with post-hoc comparisons to untreated allowing for heterscedasticity. ( D ) Quantification of nuclei per field after 5, 10, or 14 days. Data are presented as mean ± SEM ( n = 6). Two-way ANOVA with post-hoc comparisons to untreated. For all panels, P = 0.0 indicates that calculated P- values were below the limit of the program (2.2 × 10 −16 ). ns: nonsignificant.

Journal: Nucleic Acids Research

Article Title: RNA editing for the treatment of alpha-1 antitrypsin deficiency

doi: 10.1093/nar/gkaf1518

Figure Lengend Snippet: SERPINA1-994 decreases polymeric AAT accumulation in human iPSC-derived ZZ hepatocytes. ( A ) Images of cultured iPSCs after 14 days of treatment with the indicated AIMer. AAT polymer staining (green) and DAPI staining of nuclei (blue) are shown. The scale bars are 100 μm. ( B ) Quantification of polymeric AAT after 5, 10, or 14 days of treatment with increasing concentrations (0, 1.25, 2.5, 5.0, or 10 µM) of the indicated AIMer. Data are presented as mean ± SEM ( n = 6). White-adjusted two-way ANOVA with post-hoc comparisons to untreated allowing heteroscedasticity. ( C ) SERPINA1 RNA editing after 5, 10, or 14 days of treatment. Data are presented as mean ± SEM ( n = 4–6). White-adjusted two-way ANOVA with post-hoc comparisons to untreated allowing for heterscedasticity. ( D ) Quantification of nuclei per field after 5, 10, or 14 days. Data are presented as mean ± SEM ( n = 6). Two-way ANOVA with post-hoc comparisons to untreated. For all panels, P = 0.0 indicates that calculated P- values were below the limit of the program (2.2 × 10 −16 ). ns: nonsignificant.

Article Snippet: Fixed plates were immunostained with an antibody specific to the polymeric form of AAT (Hycult, Cat. No. HM2289) and quantified using a CellInsight CX7 high-content analysis platform (Thermo Fisher Scientific), and data were presented as mean fluorescence intensity per cell.

Techniques: Derivative Assay, Cell Culture, Polymer, Staining