Additional file 1: of Viral-mediated gene delivery of TMBIM6 protects the neonatal brain via disruption of NPR-CYP complex coupled with upregulation of Nrf-2 post-HI

Experimental Design. Representative figure showing experimental design and animal groups. HI, Hypoxia- Ischemia; IHC, immunohistochemistry; DHE, Dihydroethidium. (TIF 2430 kb

Additional file 9: of Viral-mediated gene delivery of TMBIM6 protects the neonatal brain via disruption of NPR-CYP complex coupled with upregulation of Nrf-2 post-HI

Statistical analysis of IL-1β and MPO positive cells at 72 h post-HI. Representative table showing detailed statistical analysis from immunohistochemistry experiments measuring the expression of IL-1β with microglia and MPO. (Data expressed as mean +/− SD; *p < 0.05 vs sham; #p < 0.05 vs vehicle; @p < 0.05 vs Ad-TMBIM6 or scramble; n = 6/group using one-way ANOVA followed by Tukey multiple-comparison post hoc analysis). (XLSX 11 kb

Additional file 7: of Viral-mediated gene delivery of TMBIM6 protects the neonatal brain via disruption of NPR-CYP complex coupled with upregulation of Nrf-2 post-HI

Statistical Analysis for Expression Levels of BI-1 Pathway Proteins at 72 h post-HI. Representative table showing detailed statistical analysis from western blot experiments measuring the levels of pathway proteins. (Data expressed as mean +/− SD; *p < 0.05 vs sham; #p < 0.05 vs vehicle; @p < 0.05 vs Ad-TMBIM6 or scramble; n = 6/group using one-way ANOVA followed by Tukey multiple-comparison post hoc analysis). (XLSX 16 kb

Additional file 6: of Viral-mediated gene delivery of TMBIM6 protects the neonatal brain via disruption of NPR-CYP complex coupled with upregulation of Nrf-2 post-HI

Statistical Analysis for Percent Infarcted Area with Intervention groups at 72 h post-HI. Representative table showing detailed statistical analysis for viral vector present in the brain and the percent infarcted area seen with different intervention groups. (Data expressed as mean +/− SD; *p < 0.05 vs sham; #p < 0.05 vs vehicle; @p < 0.05 vs Ad-TMBIM6 or scramble; n = 6–8/group using one-way ANOVA followed by Tukey multiple-comparison post hoc analysis). (XLSX 10 kb

Additional file 2: of Viral-mediated gene delivery of TMBIM6 protects the neonatal brain via disruption of NPR-CYP complex coupled with upregulation of Nrf-2 post-HI

Ad-TMBIM6 attenuates inflammation in primary microglial cells after OGD. Representative western blot bands of BI-1, P4502E1, IL-6 and IL-1β (A). Immunofluorescent staining of Iba-1 with P4502E1, BI-1 or pNrf-2 (B-D). (Green was for microglial staining, Red was for P4502E1, BI-1 or pNrf-2. Merge showed the co-localization of P4502E1, BI-1 or pNrf-2 on microglia. Scale bar 50 μm). (TIF 3861 kb

Additional file 10: of Viral-mediated gene delivery of TMBIM6 protects the neonatal brain via disruption of NPR-CYP complex coupled with upregulation of Nrf-2 post-HI

Statistical Analysis for Expression Levels of BI-1 pathway proteins in an in vitro OGD model. Representative table showing detailed statistical analysis from western blot experiments measuring the levels of BI-1, P4502E1, pNrf-2 and inflammatory markers. (Data represent +/− SD; *p < 0.05 vs control; #p < 0.05 vs vehicle; @p < 0.05 vs Ad-TMBIM6 and scramble; $p < 0.05 vs BI-1 siRNA; n = 4–5/group using one-way ANOVA followed by Tukey multiple-comparison post hoc analysis). (XLSX 14 kb

Additional file 8: of Viral-mediated gene delivery of TMBIM6 protects the neonatal brain via disruption of NPR-CYP complex coupled with upregulation of Nrf-2 post-HI

Statistical Analysis of Inflammatory Protein Expression Levels at 72 h post-HI. Representative table showing detailed statistical analysis from western blot experiments measuring the levels of pathway proteins. (Data expressed as mean +/− SD; *p < 0.05 vs sham; #p < 0.05 vs vehicle; @p < 0.05 vs Ad-TMBIM6 or scramble; n = 6/group using one-way ANOVA followed by Tukey multiple-comparison post hoc analysis). (XLSX 15 kb

Additional file 5: of Viral-mediated gene delivery of TMBIM6 protects the neonatal brain via disruption of NPR-CYP complex coupled with upregulation of Nrf-2 post-HI

Statistical Analysis for Percent Infarcted Area and Long-term studies. Representative table showing detailed statistical analysis for dose study, viral vector present in the brain and long-term neurobehavior studies. (Data expressed as mean +/− SD; *p < 0.05 vs sham; # p<0.05 vs vehicle, @ p < 0.05 vs Ad-TMBIM6 (1.6 × 1011 PFU), n = 6. One-way ANOVA followed by Tukey multiple-comparison post hoc analysis). (For long-term studies: *p<0.05 vs sham; #p<0.05 vs vehicle; n = 6–9. One-way ANOVA followed by Tukey multiple-comparison/or Holm-Sidak post hoc analysis). (XLSX 11 kb

Additional file 3: of Viral-mediated gene delivery of TMBIM6 protects the neonatal brain via disruption of NPR-CYP complex coupled with upregulation of Nrf-2 post-HI

Ad-TMBIM6 attenuates ER stress receptors’ signaling in primary microglial cells after OGD. Representative western blot bands of pIRE1α, XBP1, pPERK and peIF2α (A). Immunofluorescent staining of Iba-1 with pIRE1α or pPERK (B-C). (Green was for microglial staining, Red was for pIRE1α or pPERK. Merge showed the co-localization of pIRE1α or pPERK on microglia. Scale bar 50 μm). (TIF 2942 kb

Additional file 4: of Viral-mediated gene delivery of TMBIM6 protects the neonatal brain via disruption of NPR-CYP complex coupled with upregulation of Nrf-2 post-HI

Statistical Analysis for Endogenous Protein Expression Levels post-HI. Representative table showing detailed statistical analysis for the endogenous expression levels of BI-1, NPR, P4502E1, pNrf-2 and HO-1. (Data expressed as mean +/− SD; *p < 0.05 vs sham; # p<0.05 vs 6 h, @ p < 0.05 vs 24 h; n = 4, using one-way ANOVA followed by Tukey multiple-comparison post hoc analysis). (XLSX 12 kb