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Sfig. 1

Sfig. 1. *. *. *. *. *. *. *. C/EBP- α. p300. Dicer. Enox1. EphrinA1. MMP2. NeuroD6. RxR- γ. Gab2. FOXO1. ITG- α 4. Tubb2a. TH1-like. GAPDH. Normal SC. SCI. SCI / scramRNA. SCI / AntimiR 486. Sfig. 2. A. SCI 7 th day (Thoracic 11 of lesion). MBP /DCFDA /DAPI.

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Sfig. 1

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  1. Sfig. 1 * * * * * * * C/EBP-α p300 Dicer Enox1 EphrinA1 MMP2 NeuroD6 RxR-γ Gab2 FOXO1 ITG-α4 Tubb2a TH1-like GAPDH Normal SC SCI SCI / scramRNA SCI / AntimiR 486

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

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

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

  5. Sfig. 3 A B mmu-miR486 * RAW 264.7 / H2O2 0μM 60μM H2O2 0μM H2O2 30μM * * H2O2 60μM GAPDH 0μM 30μM 60μM H2O2 NeuroD6 C 0 30 60 RAW 264.7 / H2O2 * NeuroD6 * * * * SEPN1 TXNL1 GPx3 GAPDH H2O2 0μM 30μM 60μM SEPN1 TXNL1 GPx3

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

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

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

  9. Sfig. 7 NeuroD6/ in situ E scramRNA/SC AntimiR 486/SCI scramRNA/SC F ROS scramRNA/SCI miR 486/SC siNeuroD6/SC NeuroD6/HSP90/DAPI AntimiR 486/SCI siNeuroD6/SC rescue T10 scramRNA/NPC siNeuroD6/NPC T10 c-Myc C D In vivo SC Survival p53 DCFDA/DAPI scramRNA/SC siNeuroD6/SC scramRNA/SC siNeuroD6/SC In vitro (NPCs) p21 NeuroD6 p-PI3K B c-Myc p-Akt scramRNA/SC scramRNA/ SCI siNeuroD6/SC Bax p-Erk p53 A Caspase 3 Chaperon p-p38 * Survival/Apoptosis Cytochr C scramRNA/SC siNeuroD6/SC p-JUNK scramRNA / SC scramRNA / SCI HSP70 GAPDH miR 486 / SC p-ERK HSP90 * c-Myc GAPDH p53 HSF1 HSF2 p21

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

  11. I H Chaperon scramRNA/NPC siNeuroD6/NPC siNeuroD6/NPC scramRNA/NPC Normal NPC HSP70 HSP90 * * * GAPDH p300 HSF1 HSF2 C/EBP-α J Neurogenesis (NPCs) rescue scramRNA/NPC siNeuroD6/NPC NF160/GFAP/DAPI

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

  13. C D SCI (T10) HSF1 HSF2 SCI/ NeuroD6 MiR 486 MiR 486/ NeuroD6 siNeuroD6 siNeuroD6/NeuroD6 NF160/Cox2/DAPI * * ED1/GFAP/DAPI * * * * * * * * * * scramRNA/SC scramRNA/SC - + - - - - - - - + - - - - - - scramRNA/SC scramRNA/SC - + - - - - - - - + - - - - - - scramRNA/SCI scramRNA/SCI - - + - - - - - - - + - - - - - scramRNA/SCI scramRNA/SCI - - + - - - - - - - + - - - - - NF160/NeuroD6/DAPI miR 486/SC miR 486/SC - - - - + - - - - - - - + - - - miR486/SC miR 486/SC - - - - + - - - - - - - + - - - siNeuroD6/SC siNeuroD6/SC - - - - - - + - - - - - - - + - siNeuroD6/SC siNeuroD6/SC - - - - - - + - - - - - - - + - NeuroD6/SCI NeuroD6/SCI - - - + - + - + - - - + - + - + - - - + - + - + NeuroD6/SCI - - - + - + - + NeuroD6/SCI C/EBPα p300 /scramRNA SCI * *

  14. Sfig. 10 Mmu-miR135a-2 A - 1 2 3 8 SCI (days) - 1 2 3 8 SCI (days) Mmu-miR705 Mmu-miR135a-1 GAPDH GAPDH Normal SC Normal SC Normal SC Normal SC Normal SC Normal SC * * * SCI1day SCI1day SCI1day SCI1day SCI1day SCI1day * * - 1 2 3 8 SCI (days) * * * * * Mmu-miR135a-2 * SCI 2days SCI 2days SCI 2days SCI 2days SCI 2days SCI 2days GAPDH SCI 3days SCI 3days SCI 3days SCI 3days SCI 3days SCI 3days SCI 8days SCI 8days SCI 8days SCI 8days SCI 8days SCI 8days GAPDH miR135a-1 GAPDH miR135a-2 GAPDH miR705 - 1 2 3 8 SCI (days) - 1 2 3 8 SCI (days) Mmu-miR875 Mmu-miR873 GAPDH * * * * GAPDH Mmu-miR873 Mmu-miR875 Mmu-miR20a - 1 2 3 8 SCI (days) Mmu-miR20a * * * GAPDH * * * * * * * * GAPDH miR873 GAPDH miR875 GAPDH miR20a

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

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

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