Sfig. 1 Dicer Enox1EphrinA1 MMP2 RxR-γ NeuroD6 Gab2 FOXO1 ITG-α4Tubb2a TH1-like GAPDH SCI / AntimiR 486Normal SC SCI / scramRNASCI * * * * * C/EBP-αp300.

Παρουσίαση με θέμα: "Sfig. 1 Dicer Enox1EphrinA1 MMP2 RxR-γ NeuroD6 Gab2 FOXO1 ITG-α4Tubb2a TH1-like GAPDH SCI / AntimiR 486Normal SC SCI / scramRNASCI * * * * * C/EBP-αp300."— Μεταγράφημα παρουσίασης:

1 Sfig. 1 Dicer Enox1EphrinA1 MMP2 RxR-γ NeuroD6 Gab2 FOXO1 ITG-α4Tubb2a TH1-like GAPDH SCI / AntimiR 486Normal SC SCI / scramRNASCI * * * * * C/EBP-αp300 * *

2 Sfig. 2 MBP/DCFDA/DAPI GFAP/DCFDA/DAPI NF160/DCFDA/DAPI NeuroD6/DCFDA/DAPI A SCI 7 th day (Thoracic 11 of lesion)

3 TXNL1 DAPI SCI/AntimiR 486 7 th day (T10-T11 of lesion) SCI/scramRNA 7 th day DAPI SEPN1 NF160 DAPI GPx3 NF160 B NF160/TXNL1/DAPI NF160/GPx1/DAPI NF160/SEPN1/DAPI IgG/DAPI

4 (negative control) scram RNA/SCI AntimiR 486/SCI Normal SC MiR 486/SCSCI/antimiR 486 NF160/Cox2/DAPI SCI/scramRNA T10-T11 Normal SC MiR 486/SCSCI/antimiR 486 NF160/ED1/DAPI SCI/scramRNA T10-T11 Normal SC MiR 486/SCSCI/antimiR 486 NF160/Iba1/DAPI SCI/scramRNA T10-T11 E NF160/Cox2/DAPI NF160/ED1/DAPI NF160/Iba1/DAPI D DCFDA/DAPI SCI(T10)scramRNA/ SC MiR 486/SCmiR 486/AntimiR 486/SCI ROS(T10-11) DCFDA/DAPI B C DAPIDCFDA Iba1 DAPI DCFDA ED1 SCI 7 th day (Thoracic 10-11 of lesion) IgG/DAPI

5 mmu-miR486 0μM0μM 30μM60μM H2O2H2O2 GAPDH A * * NeuroD6 0μM0μM30μM60μMH2O2H2O2 B 0μM0μM RAW 264.7 / H 2 O 2 * NeuroD6 SEPN1 TXNL1 GPx3 GAPDH 0 30 60 SEPN1TXNL1GPx3 H 2 O 2 0μM H 2 O 2 30μMH 2 O 2 60μM C * * * * * RAW 264.7 / H 2 O 2 Sfig. 3

6 Sfig. 4 AntimiR 486 GAPDH c-Myc p53 p21 B Cell cycle - - - + - - + - scramRNA - + + + SCI A HSP90 HSP70 NeuroD6 GAPDH Chaperon AntimiR 486 - - - + - - + - scramRNA - + + + SCI

7 Sfig. 5 D HSF1 HSF2 * * miR 486/SC Normal SC scramRNA/SCI scramRNA/SC HSP90 HSP70 GAPDH scramRNA / SC Chaperon scramRNA / SCI miR 486 / SC c-Myc p53 p21 - - + AntimiR 486 - + + miR 486 In vitro (NPCs) B scramRNA + - - miR 486/NPC scramRNA/NPC C * A HSP90 HSP70 GAPDH - - + AntimiR 486 - + + miR 486 In vitro (NPCs) p-Erk p-PI3K p-Akt scramRNA + - -

8 Sfig. 6 NF160/GFAP/DAPI Tuj/GFAP/DAPI Motor neuron culture and identification C Nestin/GFAP/DAPI A2B5/GFAP/DAPI miR 486/NPC rescue NF160/GFAP/DAPI B 0 50 100 Relative positive/ DAPI (%) * * * * miR 486 / NPC rescue NF160 + GFAP + scramRNA / NPC Neurogenesis (NPCs) A scramRNA/NPC Tuj(R)/GFAP(G) Tuj/MBP/DAPINF160/GFAP/DAPINestin/GFAP/DAPI scramRNA/NPC Neurosphere scramRNA/NPC miR 486/NPC rescue

9 Sfig. 7 GAPDH HSP90 HSP70 scramRNA/SC siNeuroD6/SC Chaperon A scramRNA/SC siNeuroD6/SC Survival p-JUNK p-Akt p-p38 p-Erk p-PI3K D Cytochr C Bax Caspase 3 GAPDH p21 c-Myc p53 scramRNA/NPC siNeuroD6/NPC In vitro (NPCs) p-ERK c-Myc p53 p21 Survival/Apoptosis p53 c-Myc scramRNA/SC siNeuroD6/SC In vivo SC C NeuroD6 scramRNA / SC scramRNA / SCI miR 486 / SC B * * HSF1HSF2 siNeuroD6/SC scramRNA/SC NeuroD6/HSP90/DAPI AntimiR 486/SCI scramRNA/SC siNeuroD6/SC SCI scramRNA/ T10 scramRNA/SC miR 486/SCscramRNA/SCI AntimiR 486/SCI NeuroD6/ in situ E siNeuroD6/SC DCFDA/DAPI rescue ROS F T10

10 Sfig. 8 p-Erk PI3K scramRNA/NPC siNeuroD6/NPC Cytoch C GAPDH Bax Caspase3 p-JUNK p-p38 scramRNA/NPC siNeuroD6/NPC FG SurvivalApoptosis p-Akt eNOS iNOS GAPDH Cox2 E Inflammation scramRNA/NPC siNeuroD6/NPC Gpx3 GAPDH SEPN TXNL2 NeuroD6 D * * * scramRNA/NPC siNeuroD6/NPC Normal NPC B siNeuroD6/NPC DCFDA+ 98.5%14.0% A scramRNA/NPC siNeuroD6/NPC G0 S S Cell death scramRNA/NPC scramRNA/NPC siNeuroD6/NPC Tuj MBP NeuroD6 C Neural GAPDH GFAP GAPDH Normal NPC siNeuroD6/NPC scramRNA/NPC * * * * * TujMBP GFAP

11 scramRNA/NPC siNeuroD6/NPC GAPDH HSP90 HSP70 H Chaperon I p300 C/EBP-α HSF2HSF1 * * * siNeuroD6/NPC rescue Neurogenesis (NPCs) NF160/GFAP/DAPI J scramRNA/NPC Normal NPC siNeuroD6/NPC scramRNA/NPC

12 Sfig. 9 Nestin MAP2 * * miR 486/SC- - - - + - - - - - - - - - + -siNeuroD6/SC NeuroD6/SCI- - - + - + - + - - + - - - - -scramRNA/SCI - + - - - - - -scramRNA/SC miR 486/SC- - - - + - - - - - - - - - + -siNeuroD6/SC NeuroD6/SCI- - - + - + - + - - + - - - - -scramRNA/SCI - + - - - - - -scramRNA/SC * * * * * GFAP B * * miR 486/SC- - - - + - - - - - - - - - + -siNeuroD6/SC NeuroD6/SCI- - - + - + - + - - + - - - - -scramRNA/SCI - + - - - - - - scramRNA/SC * * scramRNA /SC NeuroD6 /SC siNeuroD6 /SC NeuroD6 GAPDH NeuroD6 /SCI A

13 HSF1 HSF2 C miR 486/SC- - - - + - - - - - - - - - + -siNeuroD6/SC NeuroD6/SCI- - - + - + - + - - + - - - - -scramRNA/SCI - + - - - - - -scramRNA/SC miR 486/SC- - - - + - - - - - - - - - + -siNeuroD6/SC NeuroD6/SCI- - - + - + - + - - + - - - - -scramRNA/SCI - + - - - - - -scramRNA/SC * * * * * * * p300C/EBPα miR 486/SC- - - - + - - - - - - - - - + -siNeuroD6/SC NeuroD6/SCI- - - + - + - + - - + - - - - -scramRNA/SCI - + - - - - - -scramRNA/SC miR486/SC- - - - + - - - - - - - - - + -siNeuroD6/SC NeuroD6/SCI - - - + - + - + - - + - - - - -scramRNA/SCI - + - - - - - -scramRNA/SC * * * * * * * SCI/ NeuroD6 NF160/Cox2/DAPI MiR 486/ NeuroD6siNeuroD6/NeuroD6 ED1/GFAP/DAPI NF160/NeuroD6/DAPI D MiR 486siNeuroD6 SCI (T10) SCI /scramRNA

14 Sfig. 10 SCI (days) - 1 2 3 8 Mmu-miR135a-1 GAPDH Normal SC SCI 1day SCI 2days SCI 3days SCI 8days GAPDH miR135a-1 * * ** * Normal SC SCI 1day SCI 2days SCI 3days SCI 8days * * * * GAPDH miR135a-2 SCI (days) - 1 2 3 8 Mmu-miR135a-2 GAPDH SCI (days) - 1 2 3 8 Mmu-miR705 GAPDH Normal SC SCI 1day SCI 2days SCI 3days SCI 8days GAPDH miR705 * * * * GAPDH miR873 Normal SC SCI 1day SCI 2days SCI 3days SCI 8days * * * * SCI (days) - 1 2 3 8 Mmu-miR873 GAPDH SCI (days) - 1 2 3 8 Mmu-miR875 GAPDH Normal SC SCI 1day SCI 2days SCI 3days SCI 8days * * * * Normal SC SCI 1day SCI 2days SCI 3days SCI 8days * * * ** GAPDH miR875 GAPDH miR20a SCI (days) - 1 2 3 8 Mmu-miR20a GAPDH Mmu-miR135a-2 Mmu-miR873 Mmu-miR875 Mmu-miR20a A

15 Sfig. 10 GAPDH Neurod6 Normal SC / scramRNA SCI / scramRNA SCI / NeuroD6 100ng SCI / NeuroD6 200ng SCI / NeuroD6 400ng * * ** - - 100 200 400 NeuroD6 (ng) NeuroD6 GAPDH SCI - + + + + * Normal SC / scramRNA SCI / scramRNA SCI / AntimiR 486 1μM SCI / AntimiR 486 3μM SCI / AntimiR 486 10μM SCI / AntimiR 486 5μM GAPDH miR486 * * ** * SCI - + + + + + AntimiR486 (μM) - - 1 3 5 10 Mmu-miR486 GAPDH * miR486 Normal SC / scramRNA Normal SC / miR 486 1μM Normal SC / miR 486 3μM Normal SC / miR 486 5μM Normal SC / miR 486 10μM mmu-miR486 (μM) - 1 3 5 10 Mmu-miR486 GAPDH * * * Neurod6 Normal SC / scramRNA Normal SC / siNeuroD6 1μM Normal SC / siNeuroD6 3μM Normal SC / siNeuroD6 5μM Normal SC / siNeuroD6 10μM NeuroD6 siNeuroD6 (μM) - 1 3 5 10 GAPDH * * * ** B

16 Sfig.11 SCI (miR 486) SCI/AntimiR 486