plvx tetone puro gfp Search Results


hipscs  (Addgene inc)
94
Addgene inc hipscs
Fig. 2 Differentiation of <t>hiPSCs</t> into neurons and analysis of neuronal markers in HS and sAD neurons. A Schematic representation of the Ngn2-mediated differentiation process used to convert hiPSCs from HS and sAD donors into neurons. Neurons were induced using doxycycline (Doxy) and selected with puromycin (Puro). B Diagram of <t>the</t> <t>lentiviral</t> constructs used in the differentiation process, including Ngn2, EGFP, and Puromycin resistance genes. C Timeline of neuronal differentiation showing key steps from hiPSC plating (DIV0), doxycycline induction (DIV1), puromycin selection (DIV3), and final analysis at DIV30. D Immunofluorescence staining of neuronal markers in HS and sAD-derived neurons at DIV30. DAPI (blue) stains nuclei, EGFP (green) marks neurons, and NEUN (red) marks mature neurons. Merged images confirm co-localization of neuronal markers. Scale bars: 50 µm. E Immunofluorescence staining of the neuronal marker MAP2 (red) in HS and sAD-derived neurons, with DAPI and EGFP labeling. F Quantification of MAP2 and NEUN median fluorescence intensity (in arbitrary units, a.u.) in HS and sAD neurons, showing no significant difference (n.s.) between groups (n = 5/group). G Representative Western blot images of MAP2 and NEUN in HS and sAD neurons, with GAPDH as the loading control. H Densitometric analysis of MAP2 and NEUN expression in HS and sAD neurons showing no significant differences in fold change(n = 5/group). Each dot represents the average of three independent experiments. Data are presented as mean ± SEM. n.s. p > 0.05, assessed by Mann Whitney Rank Sum test
Hipscs, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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hngn2 lentivirus  (Addgene inc)
93
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Fig. 2 Differentiation of <t>hiPSCs</t> into neurons and analysis of neuronal markers in HS and sAD neurons. A Schematic representation of the Ngn2-mediated differentiation process used to convert hiPSCs from HS and sAD donors into neurons. Neurons were induced using doxycycline (Doxy) and selected with puromycin (Puro). B Diagram of <t>the</t> <t>lentiviral</t> constructs used in the differentiation process, including Ngn2, EGFP, and Puromycin resistance genes. C Timeline of neuronal differentiation showing key steps from hiPSC plating (DIV0), doxycycline induction (DIV1), puromycin selection (DIV3), and final analysis at DIV30. D Immunofluorescence staining of neuronal markers in HS and sAD-derived neurons at DIV30. DAPI (blue) stains nuclei, EGFP (green) marks neurons, and NEUN (red) marks mature neurons. Merged images confirm co-localization of neuronal markers. Scale bars: 50 µm. E Immunofluorescence staining of the neuronal marker MAP2 (red) in HS and sAD-derived neurons, with DAPI and EGFP labeling. F Quantification of MAP2 and NEUN median fluorescence intensity (in arbitrary units, a.u.) in HS and sAD neurons, showing no significant difference (n.s.) between groups (n = 5/group). G Representative Western blot images of MAP2 and NEUN in HS and sAD neurons, with GAPDH as the loading control. H Densitometric analysis of MAP2 and NEUN expression in HS and sAD neurons showing no significant differences in fold change(n = 5/group). Each dot represents the average of three independent experiments. Data are presented as mean ± SEM. n.s. p > 0.05, assessed by Mann Whitney Rank Sum test
Hngn2 Lentivirus, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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plv teto hngn2 egfp puro  (Addgene inc)
96
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Fig. 2 Differentiation of <t>hiPSCs</t> into neurons and analysis of neuronal markers in HS and sAD neurons. A Schematic representation of the Ngn2-mediated differentiation process used to convert hiPSCs from HS and sAD donors into neurons. Neurons were induced using doxycycline (Doxy) and selected with puromycin (Puro). B Diagram of <t>the</t> <t>lentiviral</t> constructs used in the differentiation process, including Ngn2, EGFP, and Puromycin resistance genes. C Timeline of neuronal differentiation showing key steps from hiPSC plating (DIV0), doxycycline induction (DIV1), puromycin selection (DIV3), and final analysis at DIV30. D Immunofluorescence staining of neuronal markers in HS and sAD-derived neurons at DIV30. DAPI (blue) stains nuclei, EGFP (green) marks neurons, and NEUN (red) marks mature neurons. Merged images confirm co-localization of neuronal markers. Scale bars: 50 µm. E Immunofluorescence staining of the neuronal marker MAP2 (red) in HS and sAD-derived neurons, with DAPI and EGFP labeling. F Quantification of MAP2 and NEUN median fluorescence intensity (in arbitrary units, a.u.) in HS and sAD neurons, showing no significant difference (n.s.) between groups (n = 5/group). G Representative Western blot images of MAP2 and NEUN in HS and sAD neurons, with GAPDH as the loading control. H Densitometric analysis of MAP2 and NEUN expression in HS and sAD neurons showing no significant differences in fold change(n = 5/group). Each dot represents the average of three independent experiments. Data are presented as mean ± SEM. n.s. p > 0.05, assessed by Mann Whitney Rank Sum test
Plv Teto Hngn2 Egfp Puro, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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plvx tetone puro vector  (Addgene inc)
93
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Fig. 2 Differentiation of <t>hiPSCs</t> into neurons and analysis of neuronal markers in HS and sAD neurons. A Schematic representation of the Ngn2-mediated differentiation process used to convert hiPSCs from HS and sAD donors into neurons. Neurons were induced using doxycycline (Doxy) and selected with puromycin (Puro). B Diagram of <t>the</t> <t>lentiviral</t> constructs used in the differentiation process, including Ngn2, EGFP, and Puromycin resistance genes. C Timeline of neuronal differentiation showing key steps from hiPSC plating (DIV0), doxycycline induction (DIV1), puromycin selection (DIV3), and final analysis at DIV30. D Immunofluorescence staining of neuronal markers in HS and sAD-derived neurons at DIV30. DAPI (blue) stains nuclei, EGFP (green) marks neurons, and NEUN (red) marks mature neurons. Merged images confirm co-localization of neuronal markers. Scale bars: 50 µm. E Immunofluorescence staining of the neuronal marker MAP2 (red) in HS and sAD-derived neurons, with DAPI and EGFP labeling. F Quantification of MAP2 and NEUN median fluorescence intensity (in arbitrary units, a.u.) in HS and sAD neurons, showing no significant difference (n.s.) between groups (n = 5/group). G Representative Western blot images of MAP2 and NEUN in HS and sAD neurons, with GAPDH as the loading control. H Densitometric analysis of MAP2 and NEUN expression in HS and sAD neurons showing no significant differences in fold change(n = 5/group). Each dot represents the average of three independent experiments. Data are presented as mean ± SEM. n.s. p > 0.05, assessed by Mann Whitney Rank Sum test
Plvx Tetone Puro Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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plvx tetone puro vector  (New England Biolabs)
99
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Fig. 2 Differentiation of <t>hiPSCs</t> into neurons and analysis of neuronal markers in HS and sAD neurons. A Schematic representation of the Ngn2-mediated differentiation process used to convert hiPSCs from HS and sAD donors into neurons. Neurons were induced using doxycycline (Doxy) and selected with puromycin (Puro). B Diagram of <t>the</t> <t>lentiviral</t> constructs used in the differentiation process, including Ngn2, EGFP, and Puromycin resistance genes. C Timeline of neuronal differentiation showing key steps from hiPSC plating (DIV0), doxycycline induction (DIV1), puromycin selection (DIV3), and final analysis at DIV30. D Immunofluorescence staining of neuronal markers in HS and sAD-derived neurons at DIV30. DAPI (blue) stains nuclei, EGFP (green) marks neurons, and NEUN (red) marks mature neurons. Merged images confirm co-localization of neuronal markers. Scale bars: 50 µm. E Immunofluorescence staining of the neuronal marker MAP2 (red) in HS and sAD-derived neurons, with DAPI and EGFP labeling. F Quantification of MAP2 and NEUN median fluorescence intensity (in arbitrary units, a.u.) in HS and sAD neurons, showing no significant difference (n.s.) between groups (n = 5/group). G Representative Western blot images of MAP2 and NEUN in HS and sAD neurons, with GAPDH as the loading control. H Densitometric analysis of MAP2 and NEUN expression in HS and sAD neurons showing no significant differences in fold change(n = 5/group). Each dot represents the average of three independent experiments. Data are presented as mean ± SEM. n.s. p > 0.05, assessed by Mann Whitney Rank Sum test
Plvx Tetone Puro Vector, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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plvx tetone puro p21  (Addgene inc)
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Fig. 2 Differentiation of <t>hiPSCs</t> into neurons and analysis of neuronal markers in HS and sAD neurons. A Schematic representation of the Ngn2-mediated differentiation process used to convert hiPSCs from HS and sAD donors into neurons. Neurons were induced using doxycycline (Doxy) and selected with puromycin (Puro). B Diagram of <t>the</t> <t>lentiviral</t> constructs used in the differentiation process, including Ngn2, EGFP, and Puromycin resistance genes. C Timeline of neuronal differentiation showing key steps from hiPSC plating (DIV0), doxycycline induction (DIV1), puromycin selection (DIV3), and final analysis at DIV30. D Immunofluorescence staining of neuronal markers in HS and sAD-derived neurons at DIV30. DAPI (blue) stains nuclei, EGFP (green) marks neurons, and NEUN (red) marks mature neurons. Merged images confirm co-localization of neuronal markers. Scale bars: 50 µm. E Immunofluorescence staining of the neuronal marker MAP2 (red) in HS and sAD-derived neurons, with DAPI and EGFP labeling. F Quantification of MAP2 and NEUN median fluorescence intensity (in arbitrary units, a.u.) in HS and sAD neurons, showing no significant difference (n.s.) between groups (n = 5/group). G Representative Western blot images of MAP2 and NEUN in HS and sAD neurons, with GAPDH as the loading control. H Densitometric analysis of MAP2 and NEUN expression in HS and sAD neurons showing no significant differences in fold change(n = 5/group). Each dot represents the average of three independent experiments. Data are presented as mean ± SEM. n.s. p > 0.05, assessed by Mann Whitney Rank Sum test
Plvx Tetone Puro P21, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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plvx tetone puro lentiviral plasmids  (Addgene inc)
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Fig. 2 Differentiation of <t>hiPSCs</t> into neurons and analysis of neuronal markers in HS and sAD neurons. A Schematic representation of the Ngn2-mediated differentiation process used to convert hiPSCs from HS and sAD donors into neurons. Neurons were induced using doxycycline (Doxy) and selected with puromycin (Puro). B Diagram of <t>the</t> <t>lentiviral</t> constructs used in the differentiation process, including Ngn2, EGFP, and Puromycin resistance genes. C Timeline of neuronal differentiation showing key steps from hiPSC plating (DIV0), doxycycline induction (DIV1), puromycin selection (DIV3), and final analysis at DIV30. D Immunofluorescence staining of neuronal markers in HS and sAD-derived neurons at DIV30. DAPI (blue) stains nuclei, EGFP (green) marks neurons, and NEUN (red) marks mature neurons. Merged images confirm co-localization of neuronal markers. Scale bars: 50 µm. E Immunofluorescence staining of the neuronal marker MAP2 (red) in HS and sAD-derived neurons, with DAPI and EGFP labeling. F Quantification of MAP2 and NEUN median fluorescence intensity (in arbitrary units, a.u.) in HS and sAD neurons, showing no significant difference (n.s.) between groups (n = 5/group). G Representative Western blot images of MAP2 and NEUN in HS and sAD neurons, with GAPDH as the loading control. H Densitometric analysis of MAP2 and NEUN expression in HS and sAD neurons showing no significant differences in fold change(n = 5/group). Each dot represents the average of three independent experiments. Data are presented as mean ± SEM. n.s. p > 0.05, assessed by Mann Whitney Rank Sum test
Plvx Tetone Puro Lentiviral Plasmids, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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pb09 tre ef1a beta globin mirctrl gfp teto puro  (Addgene inc)
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Fig. 2 Differentiation of <t>hiPSCs</t> into neurons and analysis of neuronal markers in HS and sAD neurons. A Schematic representation of the Ngn2-mediated differentiation process used to convert hiPSCs from HS and sAD donors into neurons. Neurons were induced using doxycycline (Doxy) and selected with puromycin (Puro). B Diagram of <t>the</t> <t>lentiviral</t> constructs used in the differentiation process, including Ngn2, EGFP, and Puromycin resistance genes. C Timeline of neuronal differentiation showing key steps from hiPSC plating (DIV0), doxycycline induction (DIV1), puromycin selection (DIV3), and final analysis at DIV30. D Immunofluorescence staining of neuronal markers in HS and sAD-derived neurons at DIV30. DAPI (blue) stains nuclei, EGFP (green) marks neurons, and NEUN (red) marks mature neurons. Merged images confirm co-localization of neuronal markers. Scale bars: 50 µm. E Immunofluorescence staining of the neuronal marker MAP2 (red) in HS and sAD-derived neurons, with DAPI and EGFP labeling. F Quantification of MAP2 and NEUN median fluorescence intensity (in arbitrary units, a.u.) in HS and sAD neurons, showing no significant difference (n.s.) between groups (n = 5/group). G Representative Western blot images of MAP2 and NEUN in HS and sAD neurons, with GAPDH as the loading control. H Densitometric analysis of MAP2 and NEUN expression in HS and sAD neurons showing no significant differences in fold change(n = 5/group). Each dot represents the average of three independent experiments. Data are presented as mean ± SEM. n.s. p > 0.05, assessed by Mann Whitney Rank Sum test
Pb09 Tre Ef1a Beta Globin Mirctrl Gfp Teto Puro, supplied by Addgene inc, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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plvx tetone puro backbone  (New England Biolabs)
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Fig. 2 Differentiation of <t>hiPSCs</t> into neurons and analysis of neuronal markers in HS and sAD neurons. A Schematic representation of the Ngn2-mediated differentiation process used to convert hiPSCs from HS and sAD donors into neurons. Neurons were induced using doxycycline (Doxy) and selected with puromycin (Puro). B Diagram of <t>the</t> <t>lentiviral</t> constructs used in the differentiation process, including Ngn2, EGFP, and Puromycin resistance genes. C Timeline of neuronal differentiation showing key steps from hiPSC plating (DIV0), doxycycline induction (DIV1), puromycin selection (DIV3), and final analysis at DIV30. D Immunofluorescence staining of neuronal markers in HS and sAD-derived neurons at DIV30. DAPI (blue) stains nuclei, EGFP (green) marks neurons, and NEUN (red) marks mature neurons. Merged images confirm co-localization of neuronal markers. Scale bars: 50 µm. E Immunofluorescence staining of the neuronal marker MAP2 (red) in HS and sAD-derived neurons, with DAPI and EGFP labeling. F Quantification of MAP2 and NEUN median fluorescence intensity (in arbitrary units, a.u.) in HS and sAD neurons, showing no significant difference (n.s.) between groups (n = 5/group). G Representative Western blot images of MAP2 and NEUN in HS and sAD neurons, with GAPDH as the loading control. H Densitometric analysis of MAP2 and NEUN expression in HS and sAD neurons showing no significant differences in fold change(n = 5/group). Each dot represents the average of three independent experiments. Data are presented as mean ± SEM. n.s. p > 0.05, assessed by Mann Whitney Rank Sum test
Plvx Tetone Puro Backbone, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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work n a plvx tetone puro vector takara 631847 pcdna3 1  (Addgene inc)
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Fig. 2 Differentiation of <t>hiPSCs</t> into neurons and analysis of neuronal markers in HS and sAD neurons. A Schematic representation of the Ngn2-mediated differentiation process used to convert hiPSCs from HS and sAD donors into neurons. Neurons were induced using doxycycline (Doxy) and selected with puromycin (Puro). B Diagram of <t>the</t> <t>lentiviral</t> constructs used in the differentiation process, including Ngn2, EGFP, and Puromycin resistance genes. C Timeline of neuronal differentiation showing key steps from hiPSC plating (DIV0), doxycycline induction (DIV1), puromycin selection (DIV3), and final analysis at DIV30. D Immunofluorescence staining of neuronal markers in HS and sAD-derived neurons at DIV30. DAPI (blue) stains nuclei, EGFP (green) marks neurons, and NEUN (red) marks mature neurons. Merged images confirm co-localization of neuronal markers. Scale bars: 50 µm. E Immunofluorescence staining of the neuronal marker MAP2 (red) in HS and sAD-derived neurons, with DAPI and EGFP labeling. F Quantification of MAP2 and NEUN median fluorescence intensity (in arbitrary units, a.u.) in HS and sAD neurons, showing no significant difference (n.s.) between groups (n = 5/group). G Representative Western blot images of MAP2 and NEUN in HS and sAD neurons, with GAPDH as the loading control. H Densitometric analysis of MAP2 and NEUN expression in HS and sAD neurons showing no significant differences in fold change(n = 5/group). Each dot represents the average of three independent experiments. Data are presented as mean ± SEM. n.s. p > 0.05, assessed by Mann Whitney Rank Sum test
Work N A Plvx Tetone Puro Vector Takara 631847 Pcdna3 1, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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manuscript n a plvx tetone puro youbio vt9002 plko 1 puro addgene 10878 pcdh cmv mcs ef1 puro sbi cd510b pmirfp709 n1 addgene 79989 sa mir gfp  (Addgene inc)
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Fig. 2 Differentiation of <t>hiPSCs</t> into neurons and analysis of neuronal markers in HS and sAD neurons. A Schematic representation of the Ngn2-mediated differentiation process used to convert hiPSCs from HS and sAD donors into neurons. Neurons were induced using doxycycline (Doxy) and selected with puromycin (Puro). B Diagram of <t>the</t> <t>lentiviral</t> constructs used in the differentiation process, including Ngn2, EGFP, and Puromycin resistance genes. C Timeline of neuronal differentiation showing key steps from hiPSC plating (DIV0), doxycycline induction (DIV1), puromycin selection (DIV3), and final analysis at DIV30. D Immunofluorescence staining of neuronal markers in HS and sAD-derived neurons at DIV30. DAPI (blue) stains nuclei, EGFP (green) marks neurons, and NEUN (red) marks mature neurons. Merged images confirm co-localization of neuronal markers. Scale bars: 50 µm. E Immunofluorescence staining of the neuronal marker MAP2 (red) in HS and sAD-derived neurons, with DAPI and EGFP labeling. F Quantification of MAP2 and NEUN median fluorescence intensity (in arbitrary units, a.u.) in HS and sAD neurons, showing no significant difference (n.s.) between groups (n = 5/group). G Representative Western blot images of MAP2 and NEUN in HS and sAD neurons, with GAPDH as the loading control. H Densitometric analysis of MAP2 and NEUN expression in HS and sAD neurons showing no significant differences in fold change(n = 5/group). Each dot represents the average of three independent experiments. Data are presented as mean ± SEM. n.s. p > 0.05, assessed by Mann Whitney Rank Sum test
Manuscript N A Plvx Tetone Puro Youbio Vt9002 Plko 1 Puro Addgene 10878 Pcdh Cmv Mcs Ef1 Puro Sbi Cd510b Pmirfp709 N1 Addgene 79989 Sa Mir Gfp, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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plvx teton izumo1 gfp  (Addgene inc)
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Fig. 2 Differentiation of <t>hiPSCs</t> into neurons and analysis of neuronal markers in HS and sAD neurons. A Schematic representation of the Ngn2-mediated differentiation process used to convert hiPSCs from HS and sAD donors into neurons. Neurons were induced using doxycycline (Doxy) and selected with puromycin (Puro). B Diagram of <t>the</t> <t>lentiviral</t> constructs used in the differentiation process, including Ngn2, EGFP, and Puromycin resistance genes. C Timeline of neuronal differentiation showing key steps from hiPSC plating (DIV0), doxycycline induction (DIV1), puromycin selection (DIV3), and final analysis at DIV30. D Immunofluorescence staining of neuronal markers in HS and sAD-derived neurons at DIV30. DAPI (blue) stains nuclei, EGFP (green) marks neurons, and NEUN (red) marks mature neurons. Merged images confirm co-localization of neuronal markers. Scale bars: 50 µm. E Immunofluorescence staining of the neuronal marker MAP2 (red) in HS and sAD-derived neurons, with DAPI and EGFP labeling. F Quantification of MAP2 and NEUN median fluorescence intensity (in arbitrary units, a.u.) in HS and sAD neurons, showing no significant difference (n.s.) between groups (n = 5/group). G Representative Western blot images of MAP2 and NEUN in HS and sAD neurons, with GAPDH as the loading control. H Densitometric analysis of MAP2 and NEUN expression in HS and sAD neurons showing no significant differences in fold change(n = 5/group). Each dot represents the average of three independent experiments. Data are presented as mean ± SEM. n.s. p > 0.05, assessed by Mann Whitney Rank Sum test
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Fig. 2 Differentiation of hiPSCs into neurons and analysis of neuronal markers in HS and sAD neurons. A Schematic representation of the Ngn2-mediated differentiation process used to convert hiPSCs from HS and sAD donors into neurons. Neurons were induced using doxycycline (Doxy) and selected with puromycin (Puro). B Diagram of the lentiviral constructs used in the differentiation process, including Ngn2, EGFP, and Puromycin resistance genes. C Timeline of neuronal differentiation showing key steps from hiPSC plating (DIV0), doxycycline induction (DIV1), puromycin selection (DIV3), and final analysis at DIV30. D Immunofluorescence staining of neuronal markers in HS and sAD-derived neurons at DIV30. DAPI (blue) stains nuclei, EGFP (green) marks neurons, and NEUN (red) marks mature neurons. Merged images confirm co-localization of neuronal markers. Scale bars: 50 µm. E Immunofluorescence staining of the neuronal marker MAP2 (red) in HS and sAD-derived neurons, with DAPI and EGFP labeling. F Quantification of MAP2 and NEUN median fluorescence intensity (in arbitrary units, a.u.) in HS and sAD neurons, showing no significant difference (n.s.) between groups (n = 5/group). G Representative Western blot images of MAP2 and NEUN in HS and sAD neurons, with GAPDH as the loading control. H Densitometric analysis of MAP2 and NEUN expression in HS and sAD neurons showing no significant differences in fold change(n = 5/group). Each dot represents the average of three independent experiments. Data are presented as mean ± SEM. n.s. p > 0.05, assessed by Mann Whitney Rank Sum test

Journal: Alzheimer's research & therapy

Article Title: Structural and functional alterations of neurons derived from sporadic Alzheimer's disease hiPSCs are associated with downregulation of the LIMK1-cofilin axis.

doi: 10.1186/s13195-024-01632-3

Figure Lengend Snippet: Fig. 2 Differentiation of hiPSCs into neurons and analysis of neuronal markers in HS and sAD neurons. A Schematic representation of the Ngn2-mediated differentiation process used to convert hiPSCs from HS and sAD donors into neurons. Neurons were induced using doxycycline (Doxy) and selected with puromycin (Puro). B Diagram of the lentiviral constructs used in the differentiation process, including Ngn2, EGFP, and Puromycin resistance genes. C Timeline of neuronal differentiation showing key steps from hiPSC plating (DIV0), doxycycline induction (DIV1), puromycin selection (DIV3), and final analysis at DIV30. D Immunofluorescence staining of neuronal markers in HS and sAD-derived neurons at DIV30. DAPI (blue) stains nuclei, EGFP (green) marks neurons, and NEUN (red) marks mature neurons. Merged images confirm co-localization of neuronal markers. Scale bars: 50 µm. E Immunofluorescence staining of the neuronal marker MAP2 (red) in HS and sAD-derived neurons, with DAPI and EGFP labeling. F Quantification of MAP2 and NEUN median fluorescence intensity (in arbitrary units, a.u.) in HS and sAD neurons, showing no significant difference (n.s.) between groups (n = 5/group). G Representative Western blot images of MAP2 and NEUN in HS and sAD neurons, with GAPDH as the loading control. H Densitometric analysis of MAP2 and NEUN expression in HS and sAD neurons showing no significant differences in fold change(n = 5/group). Each dot represents the average of three independent experiments. Data are presented as mean ± SEM. n.s. p > 0.05, assessed by Mann Whitney Rank Sum test

Article Snippet: The lentiviral constructs used for neuronal induction of hIPSCs included: pLV-TetO-hNGN2-eGFP-Puro (a gift from Kristen Brennand; Addgene plasmid #79823), FUW-M2rtTA (a gift from Rudolf Jaenisch; Addgene plasmid #20342), pRSV-Rev, pMD2.G, pMDLg/pRRE (gifts from Didier Trono; Addgene plasmid #12253, Addgene plasmid #12259, Addgene plasmid #12251, respectively).

Techniques: Construct, Selection, Immunofluorescence, Staining, Derivative Assay, Marker, Labeling, Fluorescence, Western Blot, Control, Expressing, MANN-WHITNEY

Fig. 3 Neurons obtained from sAD-hiPSCs recapitulate the typical hallmarks of AD. A Immunofluorescence staining for Aβ using 6E10 antibody (red), DAPI (nuclei, blue), and EGFP (neurons, green) in HS and sAD hiPSC-derived neurons. sAD neurons show strong 6E10 staining, indicating the presence of Aβ pathology. Scale bars: 50 µm. B Quantification of 6E10 median fluorescence intensity (in arbitrary units, a.u.) showing a significant increase in Aβ accumulation in sAD-hNs neurons compared to HS-hNs; (n = 5/group). C Representative dot blot images of Aβ levels in DIV30 HS- and AD-hNs lysates; (D) Bar graphs showing the band intensity analysis of Aβ in HS- and sAD-hNs. Red Ponceau, RP, was used as loading index and used to normalize sample; synthetic oligomeric Aβ42 (200 nM) was used as positive control; (E) Representative WB images of p-TAU (Thr181, Ser199, Thr205, Thr217 and Thr231) levels in DIV30 HS- and sAD-hNs lysates; (F) Densitometric analysis of p-TAU (Thr181, Ser199, Thr205, Thr217 and Thr231), normalized to total human TAU (Ht7) protein levels; n = 5/group. GAPDH was used as a loading control;; norm. normalized protein levels. Each dot represents the average of three independent experiments conducted on n = 5/group. Data are presented as mean ± SEM, n.s. p > 0.05, ** p < 0.01 vs HS-hNs, assessed by Mann Whitney Rank Sum test

Journal: Alzheimer's research & therapy

Article Title: Structural and functional alterations of neurons derived from sporadic Alzheimer's disease hiPSCs are associated with downregulation of the LIMK1-cofilin axis.

doi: 10.1186/s13195-024-01632-3

Figure Lengend Snippet: Fig. 3 Neurons obtained from sAD-hiPSCs recapitulate the typical hallmarks of AD. A Immunofluorescence staining for Aβ using 6E10 antibody (red), DAPI (nuclei, blue), and EGFP (neurons, green) in HS and sAD hiPSC-derived neurons. sAD neurons show strong 6E10 staining, indicating the presence of Aβ pathology. Scale bars: 50 µm. B Quantification of 6E10 median fluorescence intensity (in arbitrary units, a.u.) showing a significant increase in Aβ accumulation in sAD-hNs neurons compared to HS-hNs; (n = 5/group). C Representative dot blot images of Aβ levels in DIV30 HS- and AD-hNs lysates; (D) Bar graphs showing the band intensity analysis of Aβ in HS- and sAD-hNs. Red Ponceau, RP, was used as loading index and used to normalize sample; synthetic oligomeric Aβ42 (200 nM) was used as positive control; (E) Representative WB images of p-TAU (Thr181, Ser199, Thr205, Thr217 and Thr231) levels in DIV30 HS- and sAD-hNs lysates; (F) Densitometric analysis of p-TAU (Thr181, Ser199, Thr205, Thr217 and Thr231), normalized to total human TAU (Ht7) protein levels; n = 5/group. GAPDH was used as a loading control;; norm. normalized protein levels. Each dot represents the average of three independent experiments conducted on n = 5/group. Data are presented as mean ± SEM, n.s. p > 0.05, ** p < 0.01 vs HS-hNs, assessed by Mann Whitney Rank Sum test

Article Snippet: The lentiviral constructs used for neuronal induction of hIPSCs included: pLV-TetO-hNGN2-eGFP-Puro (a gift from Kristen Brennand; Addgene plasmid #79823), FUW-M2rtTA (a gift from Rudolf Jaenisch; Addgene plasmid #20342), pRSV-Rev, pMD2.G, pMDLg/pRRE (gifts from Didier Trono; Addgene plasmid #12253, Addgene plasmid #12259, Addgene plasmid #12251, respectively).

Techniques: Immunofluorescence, Staining, Derivative Assay, Fluorescence, Dot Blot, Positive Control, Control, MANN-WHITNEY

Fig. 6 Neurons obtained from sAD-hiPSCs show impairments in synaptic protein expression. A Representative WB images of pre- and postsynaptic proteins in DIV30 HS- and sAD-hNs lysates; Densitometric analyses of presynaptic proteins: SYN-1, SYP and postsynaptic proteins: PSD95,GluA1; GAPDH was used as loading control; n = 5/group. C Immunofluorescence staining of synaptic markers SYN-1, SYP, PSD95, and GluA1 in HS and sAD neurons. Co-localization with EGFP (neuronal marker) is shown. sAD neurons display decreased synaptic marker expression. Scale bars: 50 µm. D Quantification of median fluorescence intensity (MFI) in HS and sAD neurons shows a significant reduction in SYN-1, SYP, PSD95, and GluA1 in sAD neurons compared to HS neurons; n = 5/group. Each dot represents the average of three independent experiments conducted on n = 5/group. Data are presented as mean ± SEM, * p < 0.05, ** p < 0.01 vs HS-hNs, assessed by Mann Whitney Rank Sum test

Journal: Alzheimer's research & therapy

Article Title: Structural and functional alterations of neurons derived from sporadic Alzheimer's disease hiPSCs are associated with downregulation of the LIMK1-cofilin axis.

doi: 10.1186/s13195-024-01632-3

Figure Lengend Snippet: Fig. 6 Neurons obtained from sAD-hiPSCs show impairments in synaptic protein expression. A Representative WB images of pre- and postsynaptic proteins in DIV30 HS- and sAD-hNs lysates; Densitometric analyses of presynaptic proteins: SYN-1, SYP and postsynaptic proteins: PSD95,GluA1; GAPDH was used as loading control; n = 5/group. C Immunofluorescence staining of synaptic markers SYN-1, SYP, PSD95, and GluA1 in HS and sAD neurons. Co-localization with EGFP (neuronal marker) is shown. sAD neurons display decreased synaptic marker expression. Scale bars: 50 µm. D Quantification of median fluorescence intensity (MFI) in HS and sAD neurons shows a significant reduction in SYN-1, SYP, PSD95, and GluA1 in sAD neurons compared to HS neurons; n = 5/group. Each dot represents the average of three independent experiments conducted on n = 5/group. Data are presented as mean ± SEM, * p < 0.05, ** p < 0.01 vs HS-hNs, assessed by Mann Whitney Rank Sum test

Article Snippet: The lentiviral constructs used for neuronal induction of hIPSCs included: pLV-TetO-hNGN2-eGFP-Puro (a gift from Kristen Brennand; Addgene plasmid #79823), FUW-M2rtTA (a gift from Rudolf Jaenisch; Addgene plasmid #20342), pRSV-Rev, pMD2.G, pMDLg/pRRE (gifts from Didier Trono; Addgene plasmid #12253, Addgene plasmid #12259, Addgene plasmid #12251, respectively).

Techniques: Expressing, Control, Immunofluorescence, Staining, Marker, Fluorescence, MANN-WHITNEY