A GCL NBL ED 18 5 PND 6
A GCL NBL ED 18. 5 PND 6 PND 3 GCL B INL PND 60 ONL
A GCL INL ONL GS CDH 11 Merge 40 x oil INL Cralbp CDH 11 Merge B INL 160 k. Da CDH 11 Merge GCL C INL ONL Chx-10 CDH 11 INL D INL HPC-1 CDH 11 ONL
ED 18. 5 PND 3 PND 6 PND 15 PND 60 A GCL INL ONL B GCL INL ONL C GCL INL ONL
Cdh 11+/+; TAg+/- A Cdh 11 -/-; TAg+/- Average Retinal Area and TAg positive cells per Eye at PND 8 140 E 5 K-W Test, p=0. 83 K-W Test, p=0. 01 n=5 120 E 5 100 E 5 9000 8000 7000 6000 5000 4000 3000 2000 1000 80 E 5 60 E 5 40 E 5 20 E 5 Cdh 11+/+; TAg+/- C Cdh 11+/-; TAg+/- Cdh 11 -/-; TAg+/- Ratio of Total TAg-positive Cells to Total Retinal Area 0. 10 0. 09 K-W Test: P=0. 0098 0. 07 0. 06 0. 05 0. 04 0. 03 0. 02 0. 01 0. 00 Cdh 11+/+; TAg+/- Cdh 11+/-; TAg+/- Cdh 11 -/-; TAg+/- TAg +ve [cells] 160 E 5 % TAg +ve cells / retina (cells/pixels) Retinal Area [pixels] B Cdh 11+/-; TAg+/-
Additional Expts Done: P 28 -PCNA Cdh 11+/+; TAg+/- A Cdh 11+/-; TAg+/- Cdh 11 -/-; TAg+/- Percent Tumor Area per Retinal Area at PND 28 B % Total Tumor Area per Retina [pixels] 9 8 K-W Test: 0. 0155 7 6 5 4 3 2 1 0 Cdh 11+/+; TAg+/- Cdh 11+/-; TAg+/- Cdh 11 -/-; TAg+/-
Cdh 11+/+; TAg+/- tional Expts A Done: PCNA Caspase-3 Cdh 11+/-; TAg+/- Cdh 11 -/-; TAg+/- Average Retinal and Tumor Areas per Eye at PND 84 B 140 E 5 K-W p=0. 07 K-W p=0. 42 Retinal and Tumor Area [pixels] 120 E 5 100 E 5 80 E 5 60 E 5 40 E 5 20 E 5 0 Cdh 11+/+; TAg+/- Cdh 11+/-; TAg+/- Cdh 11 -/-; TAg+/- Percent Tumor Area per Retinal Area at PND 84 C % Total Tumor Area per Retina [pixels] 30 25 K-W Test P=0. 935 20 15 10 5 0 Cdh 11+/+; TAg+/- Cdh 11+/-; TAg+/- Cdh 11 -/-; TAg+/-
A Tumor Growth From a Single Cell at PND 8 to PND 84 1200 K-W Test P=0. 008 Fold increase in tumor growth 1000 800 600 400 200 0 Cdh 11+/+; TAg+/- Cdh 11+/-; TAg+/- Cdh 11 -/-; TAg+/- Title? K-W Test P=0. 008 1000 P 84 Tumour / P 8 Average Tag+ B 800 600 400 200 0 Cdh 11+/+; TAg+/- Cdh 11+/-; TAg+/- Cdh 11 -/-; TAg+/-
A Cdh 11+/+ Chx-10 160 k. Da HPC-1 Cralbp Brd-U Cadherin-2 Cdh 11 -/- B Cdh 11+/+ Cdh 11 -/- C Cdh 11+/+ Cdh 11 -/-
TAg A B C cadherin-11
Number of Positively Stained Cells Hes 5 and Caspase-3 Positive cells in retinas of mice 80 p = 0. 35 70 60 CDH 11+/+; TAg+/- 50 CDH 11 -/-; TAg+/- 40 30 20 DON’T LIKE THESE COLORS 10 0 Hes-5 Caspase-3 Additional Expts Done: P 8: Hes 5, Caspase 3, PCNA, Cralbp PCNA Clone PC 10; Dako, Denmark
Antibody Name Company Dilution Used SV 40 Tag (Pab 101) mouse monoclonal Santa Cruz Biotechnology Cat# SC-147, Lot# A 2506, 1: 200 CDH 11 - clone CDH 113 H mouse monoclonal Gift from Dr. St. John at ICOS Corp. 1: 2500 CDH 2 (N-cadherin) mouse monoclonal BD Biosciences Pharmigen Cat# 610920, Lot# 06247 1: 2000 Brd. U (purified antibromodeoxyuridine) mouse monoclonal BD Biosciences Pharmigen Cat# 555627, Lot#52817, 1: 200 Progenitors and Bipolars: Chx 10 sheep polycolonal Gift from Rod Bremner, UHN 1: 1000 Apoptosis: activated caspase-3: anti-h/m Caspase 3 (active) rabbit polyclonal R&D Systems Cat# AF 835, Lot# CFZ 326011 1: 500 Early Müller Glia: Hes-5 rabbit polyclonal ABCAM Cat# ab 25374 1: 50 Müller Glia: CRALBP Rabbit polyclonal Vimentin goat polyclonal Gift from John Saari 1: 6000 Santa Cruz Biotechnology Cat# Lot# 1: 100 Ganglion - Brn 3 b goat polyclonal Santa Cruz Biotechnology Cat# sc-6026 1: 100 Amacrine – Syntaxin clone HPC 1 mouse monoclonal Sigma Cat# S 0664 1: 200 Horizontal – 160 k. Da mouse monoclonal Sigma 1: 40 ?
12 C Tumor growth from PND 28 - PND 84 Two-way ANOVA p=0. 0972 Fold increase tumor growth 10 8 6 4 2 0 Cdh 11+/+; TAg+/- Cdh 11+/-; TAg+/- Cdh 11 -/-; TAg+/-
Ratio of Total TAg-positive cells to total retinal area C TAg +ve cells / retina (cells/pixels) 0. 10 K-W Test: p=0. 01* 0. 09 0. 08 0. 07 0. 06 0. 05 0. 04 0. 03 0. 02 0. 01 0. 00 Cdh 11+/+; TAg+/- Cdh 11+/-; TAg+/- Cdh 11 -/-; TAg+/- Percent Total Tumor Volume in Retinas of Mice at PND 84 C % Total Tumor Volume per Retina [pixels] 30 25 K-W Test P=0. 42 20 15 10 5 0 Cdh 11+/+; TAg+/- Cdh 11+/-; TAg+/- Cdh 11 -/-; TAg+/-
Tumor growth from PND 28 - PND 84 12 C Two-way ANOVA p=0. 0972 Fold increase tumor growth 10 8 6 4 2 0 Cdh 11+/+; TAg+/- 450 Cdh 11+/-; TAg+/- Cdh 11 -/-; TAg+/- Tumor Growth from a single cell at PND 8 to PND 84 400 Fold increase tumor growth 350 300 250 200 150 100 50 0 Cdh 11+/+; TAg+/- Cdh 11+/-; TAg+/- Cdh 11 -/-; TAg+/- Tumor Growth from a single cell at PND 8 to PND 84 Fold increase tumor growth 1200 1000 800 600 400 200 0 Cdh 11+/+; TAg+/- Cdh 11+/-; TAg+/- Cdh 11 -/-; TAg+/-
160 E 5 Avg Retinal Area [pixels] 140 E 5 100 E 5 80 E 5 60 E 5 40 E 5 20 E 5 Avg Tag +ve [cells] 120 E 5 9000 8000 7000 6000 5000 4000 3000 2000 1000 Cdh 11+/+; TAg+/- Cdh 11+/-; TAg+/- Cdh 11 -/-; TAg+/-
B K-W Test p=0. 42*, K-W Test p=0. 07**
1200 1000 800 600 400 200 0
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