Journal: Scientific Reports

Article Title: Co-regulation of the Notch and Wnt signaling pathways promotes supporting cell proliferation and hair cell regeneration in mouse utricles

doi: 10.1038/srep29418

Figure Lengend Snippet: (Scale bar = 10 μm) (A) The untreated P3 mouse utricles cultured for 3 days showed very few EdU+/Sox2+ cells in the sensory epithelium. (B) When utricles were treated with DAPT for 3 days, there were some EdU+/Sox2+ cells in the sensory epithelium. (C) When utricles were treated with QS11 for 3 days, there were few EdU+/Sox2+ cells in the sensory epithelium and the number of SCs did not change significantly ( p > 0.05). (D) In the utricles treated with a combination of DAPT and QS11 for 3 days, there were more EdU+/Sox2+ cells compared to the DAPT-only and the QS11-only group ( p < 0.05). The number of SCs in the striolar region was greater than the control group ( p < 0.05). (D-1) The high magnifications of image D show two of EdU+/Sox2+ cells in the utricles. (G) The histograms show differences in the number of EdU+/Sox2+ cells between the groups cultured for 3 days. (H) The histograms show differences in the number of SCs between the groups cultured for 3 days. (E) The untreated P3 mouse utricles cultured for 14 days showed very few EdU+/Sox2+ cells in the sensory epithelium. (F) In the cultured utricles co-treated with DAPT and QS11 and supplied with 10 μM EdU, there were many EdU+/Sox2+ cells in the utricles and a large number of Sox2+ SCs. (F-1) The EdU+/Sox2+ cells in the utricles are shown at high magnification. (I) The number of EdU+/Sox2+ SCs in the 14-day culture was significantly greater than in the 3-day cultured. (J) The histograms show differences in the number of SCs between the utricles co-treated with DAPT+QS11 for 3 days and 14 days. The cells were counted per 100 μm × 100 μm in the striolar or extrastriolar region of the utricles. (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: The cultures were then treated with DAPT (γ-secretase inhibitor IX, N- [N- (3,5- difluorophenacetyl) -l-alanyl] -S-phenylglycine t-butyl ester, EMD, Gibbstown, NJ) and QS11 ((2S)-2-[2-(Indan-5-yloxy)-9-(1,1′-biphenyl-4-yl) methyl]-9H-purin-6- ylamino)-3-phenyl-propan-1-ol, Tocris Biosciences, USA), which modulates ARF-GTP levels and synergizes with the Wnt/β-catenin signaling pathway to upregulate β-catenin nuclear translocation.

Techniques: Cell Culture, Control

Journal: Scientific Reports

Article Title: Co-regulation of the Notch and Wnt signaling pathways promotes supporting cell proliferation and hair cell regeneration in mouse utricles

doi: 10.1038/srep29418

Figure Lengend Snippet: (Scale bar = 10 μm) (A) The untreated P3 mouse utricles cultured for 3 days. (B) When utricles were treated with DAPT for 3 days, there were few EdU+/Myo7a+ cells in the sensory epithelium and the number of HCs increased ( p < 0.05 vs. the control). (C) When the utricles were treated with QS11 for 3 days, there were few EdU+/Myo7a+ cells in the sensory epithelium and the number of HCs did not change significantly ( p > 0.05). (D) In the DAPT+QS11 combination-treated utricles, there were some EdU+/Myo7a+ cells in the utricles, which were mostly in the striolar region. There were more HCs than the other groups ( p < 0.05). (D-1) The high magnifications of the image in ( D ) show the EdU+/Myo7a+ cell in the utricle. (G) The histograms show differences in the number of EdU+ HCs between these groups cultured for 3 days. (H) The histograms show the differences in the number of HCs between these groups cultured for 3 days. (E) The untreated P3 mouse utricles cultured for 14 days. (F) In the utricles treated with 10 μM EdU and a combination of DAPT and QS11 for 14 days, there were some EdU+/Myo7a+ cells, which were found mostly in the striolar region. The number of HCs in the utricles had clearly increased. (F-1) The high magnifications of image F show EdU+/Myo7a+ cells in the utricles. (I) In the cultured utricles co-treated with DAPT and QS11 and supplied with 10 μM EdU for 14 days, the number of EdU+/Myo7a+ cells was significantly increased compared with the utricles co-treated for 3 days. (J) The histograms show the differences in the number of HCs between the utricles co-treated with DAPT + QS11 for 3 days or 14 days. The cells were counted per 100 μm × 100 μm in the striolar or extrastriolar region of the utricles (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: The cultures were then treated with DAPT (γ-secretase inhibitor IX, N- [N- (3,5- difluorophenacetyl) -l-alanyl] -S-phenylglycine t-butyl ester, EMD, Gibbstown, NJ) and QS11 ((2S)-2-[2-(Indan-5-yloxy)-9-(1,1′-biphenyl-4-yl) methyl]-9H-purin-6- ylamino)-3-phenyl-propan-1-ol, Tocris Biosciences, USA), which modulates ARF-GTP levels and synergizes with the Wnt/β-catenin signaling pathway to upregulate β-catenin nuclear translocation.

Techniques: Cell Culture, Control

Journal: Scientific Reports

Article Title: Co-regulation of the Notch and Wnt signaling pathways promotes supporting cell proliferation and hair cell regeneration in mouse utricles

doi: 10.1038/srep29418

Figure Lengend Snippet: (Scale bar = 10 μm) ( A ) The untreated P3 mouse utricles cultured for 7 days. ( B ) The P3 mouse utricles were treated with gentamicin for 48 hours and then cultured for another 7 days. ( C ) When utricles were treated with DAPT for 7 days after being treated with gentamicin, there were some EdU+/Sox2+ cells in the sensory epithelium. ( D ) When utricles were treated with QS11 for 7 days after being treated with gentamicin, there were few EdU+/Sox2+ cells in the sensory epithelium. ( E ) When utricles were treated with a combination of DAPT+QS11 for 7 days after being treated with gentamicin, there were more EdU+/Sox2+ cells in the sensory epithelium. ( E-1 ) The high magnifications of picture E show EdU+/Sox2+ cells in the utricles. ( H ) The histograms show differences in the number of EdU+/Sox2+ cells between these groups cultured for 7 days. ( I ) The histograms show differences in the number of SCs between these groups cultured for 7 days. (F) The P3 mouse utricles were treated with gentamicin for 48 hours and cultured for 14 days. The HCs were clearly damaged. (G) In the cultured utricles co-treated with DAPT and QS11 and supplied with 10 μM EdU, there were many more EdU+/Sox2+ cells in the utricles compared with the utricles co-treated for 7 days. (G-1) The high magnifications of picture G show EdU+/Sox2+ cells in the utricles. (J) The histograms show that the number of EdU+/SCs in the 14-day culture group was significantly greater than in the 7-day culture group. (K) The histograms show the differences in the number of SCs between the utricles co-treated with DAPT+QS11 for 7 days and 14 days. The cells were counted per 100 μm × 100 μm in the striolar or extrastriolar region of the utricles. (* p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The cultures were then treated with DAPT (γ-secretase inhibitor IX, N- [N- (3,5- difluorophenacetyl) -l-alanyl] -S-phenylglycine t-butyl ester, EMD, Gibbstown, NJ) and QS11 ((2S)-2-[2-(Indan-5-yloxy)-9-(1,1′-biphenyl-4-yl) methyl]-9H-purin-6- ylamino)-3-phenyl-propan-1-ol, Tocris Biosciences, USA), which modulates ARF-GTP levels and synergizes with the Wnt/β-catenin signaling pathway to upregulate β-catenin nuclear translocation.

Techniques: Cell Culture

Journal: Scientific Reports

Article Title: Co-regulation of the Notch and Wnt signaling pathways promotes supporting cell proliferation and hair cell regeneration in mouse utricles

doi: 10.1038/srep29418

Figure Lengend Snippet: (Scale bar = 10 μm) ( A ) The untreated P3 mouse utricles were cultured for 7 days. ( B ) The P3 mouse utricles were treated with gentamicin for 48 hours and cultured for 7 days. ( C ) When utricles were treated with DAPT for 7 days after HC loss, there were few EdU+/Myo7a+ cells in the utricles. ( D ) When utricles were treated with QS11 for 7 days after being treated with gentamicin, there were few EdU+/Myo7a+ cells in the utricles. ( E ) When utricles were treated with a combination of DAPT + QS11 for 7 days after being treated with gentamicin, there were some EdU+/Myo7a+ cells, and the number of HCs was greater than the other groups ( p < 0.05). ( E1 ) The high magnifications of picture E show the EdU+/Myo7a+ cell in the utricle. ( H ) The histograms show differences in the number of EdU+/HCs between these groups cultured for 7 days. ( I ) The histograms show differences in the number of HCs between these groups cultured for 7 days. (F) The P3 mouse utricles were treated with gentamicin for 48 hours and cultured for 14 days. (G) In the cultured utricles co-treated with DAPT and QS11 and supplied with 10 μM EdU for 14 days, there were some EdU+/Myo7a+ cells in the utricles. (G-1) The high magnifications of picture image G show the EdU+/Myo7a+ cells in the utricles. (J) The histograms show that the number of EdU+/Myo7a+ cells in the 14-day culture group was significantly greater than in the 7-day culture group. (K) The histograms show the differences in the number of HCs between the utricles co-treated with DAPT and QS11 for 7 days and those treated for 14 days. The cells were counted per 100 μm × 100 μm in the striolar or extrastriolar region of the utricles (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: The cultures were then treated with DAPT (γ-secretase inhibitor IX, N- [N- (3,5- difluorophenacetyl) -l-alanyl] -S-phenylglycine t-butyl ester, EMD, Gibbstown, NJ) and QS11 ((2S)-2-[2-(Indan-5-yloxy)-9-(1,1′-biphenyl-4-yl) methyl]-9H-purin-6- ylamino)-3-phenyl-propan-1-ol, Tocris Biosciences, USA), which modulates ARF-GTP levels and synergizes with the Wnt/β-catenin signaling pathway to upregulate β-catenin nuclear translocation.

Techniques: Cell Culture

Journal: Scientific Reports

Article Title: Co-regulation of the Notch and Wnt signaling pathways promotes supporting cell proliferation and hair cell regeneration in mouse utricles

doi: 10.1038/srep29418

Figure Lengend Snippet: There were some EdU+/Sox2+ cells in the DAPT and QS11 co-treated adult mouse utricles. (Scale bar = 10 μm).

Article Snippet: The cultures were then treated with DAPT (γ-secretase inhibitor IX, N- [N- (3,5- difluorophenacetyl) -l-alanyl] -S-phenylglycine t-butyl ester, EMD, Gibbstown, NJ) and QS11 ((2S)-2-[2-(Indan-5-yloxy)-9-(1,1′-biphenyl-4-yl) methyl]-9H-purin-6- ylamino)-3-phenyl-propan-1-ol, Tocris Biosciences, USA), which modulates ARF-GTP levels and synergizes with the Wnt/β-catenin signaling pathway to upregulate β-catenin nuclear translocation.

Techniques: