Fig. 1

Normal cell composition but neuronal enlargement of Tsc1-cko mouse retina. a Distribution of cells underwent Cre-mediated deletion of Tsc1 gene in E14.5 Tsc1-cko;R26^+/lacZ mouse retina was visualized indirectly by immunodetection of ß-galactosidase (ß-gal), which is expressed from a lacZ gene at Cre-recombined R26R locus. Activities of mTORC1 and mTORC2 in the retinas were also measured by immunodetection of pS6 and pAkt(S473), respectively. Scale bars, 100 μm. b Relative levels of mTOR pathway components in the mouse retinas were examined by western blotting (WB) with antibodies against corresponding proteins. SM size marker. c Hematoxylin and eosin (H&E) staining images of P14 Tsc1-het and Tsc1-cko littermate mouse retinal sections. Sizes of blue and green bars in two bottom images are same. Scale bars, 100 μm. d P14 littermate mouse eye sections were stained with antibodies that recognize Brn3b (RGC), Pax6 (AC), Calbindin (AC subset and HZ [arrowheads]), Chx10 (BP), Rhodopsin (Rhod; rPR), green/red-opsin (G/R-opsin; cPR), and Sox9 (MG). Scale bars, 200 μm. e Relative numbers of cells expressing the markers in the Tsc1-cko retinas were obtained by comparing with those in the Tsc1-het retinas. Numbers of retina analyzed are 4 (from 3 independent litters). f HZ, rod BP, and AC cells in P14 Tsc1-het and Tsc1-cko littermate mouse retinas are visualized by immunostainings with antibodies detecting respective markers Calbindin, protein kinase C-α (PKCα), and Syntaxin. Arrowheads indicate cell bodies of those retinal neurons. g Average area of the neuronal cell body in P14 Tsc1-cko mouse retinas was compared with that of littermate Tsc1-het mouse retinas. Values are averages of 200 cells in 4 different mouse retinas collected from 3 independent litters. h (Left) P14 Tsc1-het and Tsc1-cko mouse retinal cells were analyzed by FACS to compare their relative cell sizes by measuring forward scatter (FSC) values. (Right) Relative sizes of Chx10::GFP-positive bipolar cells in those littermate mouse retinas were also compared by measuring FSC values. i Mean sizes of the cells in the Tsc1-cko mouse retinas were obtained and shown in a graph as relative values to Tsc1-het samples (n = 3). Error bars in the graphs denote standard deviations (SD). P-values are obtained by Student’s t-test and shown in the graphs (*<0.05; **<0.01; ***<0.001)