Supplementary Material 2

Identification of significant canonical pathways by IPA. The top 5 pathways at

each time point of hypoxia exposure (3, 6 and 12 h) for both hypoxia and NAP

supplemented hypoxia (NAP + Hypoxia) are presented. The intensity of node

colour represents up (red), down (green) or no regulation (no colour).

S.No. 3h hypoxia S.No. 3h hypoxia + NAP

1 Mitochondrial dysfunction 1 Glycolysis I

2 RhoGDI signaling 2 NRF2-mediated oxidative stress response

3 NADH Repair 3 NADH Repair

4 Glycerol-3-phosphate Shuttle 4 Superoxide Radicals Degradation

5 Superoxide Radicals Degradation 5 nNOS signaling in neurons

Table 1: IPA identified significant canonical pathways altered during 3 h hypoxia exposure and NAP supplementation in rat brain hippocampus. Pathways are arranged in order to their P-value.

Figure 1: Ingenuity Pathway Analysis (IPA) of hypoxia (3h) regulated canonical pathway “Mitochondrial dysfunction” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 2: Ingenuity Pathway Analysis (IPA) of hypoxia (3h) regulated canonical pathway “RhoGDI signaling” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 3: Ingenuity Pathway Analysis (IPA) of hypoxia (3h) regulated canonical pathway “NADH Repair” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 4: Ingenuity Pathway Analysis (IPA) of hypoxia (3h) regulated canonical pathway “Glycerol-3-phosphate shuttle” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 5: Ingenuity Pathway Analysis (IPA) of hypoxia (3h) regulated canonical pathway “Superoxide Radicals Degradation” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 6: Ingenuity Pathway Analysis (IPA) of hypoxia (3h) regulated canonical pathway “Glycolysis I” in NAP supplemented brain hippocampus exposed to hypoxia (NAP + Hypoxia). Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 7: Ingenuity Pathway Analysis (IPA) of hypoxia (3h with NAP supplementation) regulated canonical pathway “NRF2-mediated oxidative stress response” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 8: Ingenuity Pathway Analysis (IPA) of hypoxia (3h with NAP supplementation) regulated canonical pathway “NADH Repair” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 9: Ingenuity Pathway Analysis (IPA) of hypoxia (3h with NAP supplementation) regulated canonical pathway “Superoxide Radicals Degradation” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 10: Ingenuity Pathway Analysis (IPA) of hypoxia (3h with NAP supplementation) regulated canonical pathway “nNOS signaling in neurons” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Table 2: IPA identified significant canonical pathways altered during 6 h hypoxia exposure and NAP supplementation in rat brain hippocampus. Pathways are arranged in order to their p value

S.No. 6h hypoxia S.No. 6h hypoxia + NAP

1 Mitochondrial dysfunction 1 NRF2-mediated oxidative stress response

2 NRF2-mediated oxidative stress response

2 Glutamine biosynthesis

3 Glycolysis I 3 NADH Repair

4 NADH Repair 4 Superoxide Radicals Degradation

5 Superoxide Radicals Degradation 5 Acyl-CoA hydrolysis

Figure 11: Ingenuity Pathway Analysis (IPA) of hypoxia (6h) regulated canonical pathway “Mitochondrial dysfunction” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 12: Ingenuity Pathway Analysis (IPA) Analysis of hypoxia (6h) regulated canonical pathway “NRF2-mediated oxidative stress response” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 13: Ingenuity Pathway Analysis (IPA) Analysis of hypoxia (6h) regulated canonical pathway “Glycolysis I” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 14: Ingenuity Pathway Analysis (IPA) of hypoxia (6h) regulated canonical pathway “NADH Repair” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 15: Ingenuity Pathway Analysis (IPA) of hypoxia (6h) regulated canonical pathway “Superoxide Radicals Degradation” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 16: Ingenuity Pathway Analysis (IPA) of hypoxia (6h with NAP supplementation) regulated canonical pathway “NRF2-mediated oxidative stress response” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 17: Ingenuity Pathway Analysis (IPA) of hypoxia (6h with NAP supplementation) regulated canonical pathway “Glutathione-mediated detoxification” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 18: Ingenuity Pathway Analysis (IPA) of hypoxia (6h with NAP supplementation) regulated canonical pathway “NADH Repair” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 19: Ingenuity Pathway Analysis (IPA) of hypoxia (6h with NAP supplementation) regulated canonical pathway “Superoxide Radicals Degradation” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 20: Ingenuity Pathway Analysis (IPA) of hypoxia (6h with NAP supplementation) regulated canonical pathway “Acyl-CoA hydrolysis” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Table 3: IPA identified significant canonical pathways altered during 12 h hypoxia exposure and NAP supplementation in rat brain hippocampus. Pathways are arranged in order to their P-value.

S.No. 12h hypoxia S.No. 12h hypoxia + NAP

1 Mitochondrial dysfunction 1 NRF2-mediated oxidative stress response

2 Gluconeogenesis I 2 Superoxide Radicals Degradation

3 Glycolysis I 3 Signaling by Rho family GTPases

4 Glutathione-mediated detoxification 4 AMPK signaling

5 Glycerol-3-phosphate Shuttle 5 Glutathione-mediated detoxification

Figure 21: Ingenuity Pathway Analysis (IPA) of hypoxia (12h) regulated canonical pathway “Mitochondrial dysfunction” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 22: Ingenuity Pathway Analysis (IPA) of hypoxia (12h) regulated canonical pathway “Gluconeogenesis I” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 23: Ingenuity Pathway Analysis (IPA) of hypoxia (12h) regulated canonical pathway “Glycolysis I” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 24: Ingenuity Pathway Analysis (IPA) of hypoxia (12h) regulated canonical pathway “Glutathione-mediated detoxification” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 25: Ingenuity Pathway Analysis (IPA) of hypoxia (12h) regulated canonical pathway “Glycerol-3-phosphate Shuttle” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 26: Ingenuity Pathway Analysis (IPA) of hypoxia (12h with NAP supplementation) regulated canonical pathway “NRF2-mediated oxidative stress response” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 27: Ingenuity Pathway Analysis (IPA) of hypoxia (12h with NAP supplementation) regulated canonical pathway “Superoxide Radicals Degradation” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 28: Ingenuity Pathway Analysis (IPA) of hypoxia (12h with NAP supplementation) regulated canonical pathway “Signaling by Rho family GTPases” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 29: Ingenuity Pathway Analysis (IPA) of hypoxia (12h with NAP supplementation) regulated canonical pathway “AMPK signaling” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.

Figure 30: Ingenuity Pathway Analysis (IPA) of hypoxia (12h with NAP supplementation) regulated canonical pathway “Glutathione-mediated detoxification” in brain hippocampus exposed to hypoxia. Differentially regulated proteins are shown in red and green, depicting up- and down-regulation after hypoxic exposure.