Journal: Scientific Reports

Article Title: Alpha 2 Na + ,K + -ATPase silencing induces loss of inflammatory response and ouabain protection in glial cells

doi: 10.1038/s41598-017-05075-9

Figure Lengend Snippet: α2-Na + ,K + -ATPase is silenced by siRNA. ( A ) Representative western blotting of glial cells after 72 hours of treatment with Lipofectamine RNAiMAX of α2-Na + ,K + -ATPase siRNA and a scrambled sequence. ( B ) Western blotting bands were analyzed, and the α2/β-actin ratio was calculated (n = 6). *p < 0.05 vs scrambled sequence. The full-length blots are included in the Supplementary Information file.

Article Snippet: The Na + /K + -ATPase α2 siRNA (Santa Cruz Biotechnology) was used for the experiments and the scramble sequence used was Negative Control High GC Duplex (Invitrogen).

Techniques: Western Blot, Sequencing

Journal: Scientific Reports

Article Title: Alpha 2 Na + ,K + -ATPase silencing induces loss of inflammatory response and ouabain protection in glial cells

doi: 10.1038/s41598-017-05075-9

Figure Lengend Snippet: Anti-inflammatory effect of OUA against LPS and the effect of LPS in glial cells is lost after the silencing. The cytosolic fraction of protein from glial cells (siRNA or scramble) were extracted treated with OUA (10 µM) and LPS (1 µg/mL), and western blotting was performed for pERK, tERK, RelA (p65) and β-actin (n = 5). ( A ) Representative western blotting and densitometric analysis (arbitrary units, A.U.) of pERK/tERK ratio in the scramble group, *p < 0.05 vs control, OUA and O + L. ( B ) Representative western blotting and densitometric analysis (arbitrary units, A.U.) of the pERK/tERK ratio in the siRNA group. ( C ) Representative western blotting and densitometric analysis (arbitrary units, A.U.) of the RelA (p65)/β-actin ratio in the scrambled group, *p < 0.05 vs control and O + L. ( D ) Representative western blotting and densitometric analysis (arbitrary units, A.U.) of the RelA (p65)/β-actin ratio in the siRNA group. The full-length blots are included in the Supplementary Information file.

Article Snippet: The Na + /K + -ATPase α2 siRNA (Santa Cruz Biotechnology) was used for the experiments and the scramble sequence used was Negative Control High GC Duplex (Invitrogen).

Techniques: Western Blot, Control

Journal: Scientific Reports

Article Title: Alpha 2 Na + ,K + -ATPase silencing induces loss of inflammatory response and ouabain protection in glial cells

doi: 10.1038/s41598-017-05075-9

Figure Lengend Snippet: The lack of α2-Na + ,K + -ATPase decreases TNF-α levels. Cell culture supernatants of scrambled sequence and siRNA cells after treatment with LPS (1 µg/mL) and OUA (10 µM) were collected and used to measure TNF-α levels by ELISA (n = 6). ( A ) The levels of TNF-α in the scrambled group, *p < 0.05 vs control and OUA. ( B ) The levels of TNF-α in the siRNA group, *p < 0.05 vs control and OUA. ( C ) Comparison of the levels of TNF-α in the siRNA and scramble groups, *p < 0.05 vs scramble + LPS and scramble + O + L.

Article Snippet: The Na + /K + -ATPase α2 siRNA (Santa Cruz Biotechnology) was used for the experiments and the scramble sequence used was Negative Control High GC Duplex (Invitrogen).

Techniques: Cell Culture, Sequencing, Enzyme-linked Immunosorbent Assay, Control, Comparison

Journal: Scientific Reports

Article Title: Alpha 2 Na + ,K + -ATPase silencing induces loss of inflammatory response and ouabain protection in glial cells

doi: 10.1038/s41598-017-05075-9

Figure Lengend Snippet: α2-Na + ,K + -ATPase is not related to changes in IL-10 levels. Cell culture supernatants of scrambled sequence and siRNA cells after treatment with LPS(1 µg/mL) and OUA (10 µM) were collected and used to measure IL-1β and IL-10 levels by ELISA (n = 6). ( A ) The levels of IL-1β in the scrambled group, *p < 0.05 vs control. ( B ) The levels of IL-1β in the siRNA group showed no difference between the groups. ( C ) The levels of IL-10 in the scramble group, *p < 0.05 vs control, OUA and O + L. ( D ) The levels of IL-10 in the siRNA group, *p < 0.05 vs control, OUA and O + L.

Article Snippet: The Na + /K + -ATPase α2 siRNA (Santa Cruz Biotechnology) was used for the experiments and the scramble sequence used was Negative Control High GC Duplex (Invitrogen).

Techniques: Cell Culture, Sequencing, Enzyme-linked Immunosorbent Assay, Control