Fig 1: Characterization of the P0CreIAbfl/fl mouse line. (a) P0Cre mice were crossed with IAbfl/fl mice to generate homozygous P0CreIAbfl/fl mice. Wildtype IAbfl/fl mice served as controls in all experiments. Acknowledgements for drawings of mice to Prof. Dr. Sven Meuth and Heike Blum, Institute for Translational Neurology, University Hospital Münster, Münster, Germany. (b) One week after chronic constriction injury, longitudinal paraffin sections (thickness 6 µm) of the sciatic nerve from wildtype IAbfl/fl mice (top panels) and P0CreIAbfl/fl mice (bottom panels) were stained against S100 (green signal), MHC class II (MHC-II; red signal) using fluorescently labelled secondary antibodies; nuclei were stained with DAPI. White arrow indicates an MHC-II expressing macrophage. Right panels depict higher magnifications of the areas indicated in the left panels.
Fig 2: Effect of safflor yellow B (SYB) on target protein expression in RAECsRAECs were used to establish OGD model. Total proteins in RAECs from different groups were extracted and measured. Thirty micrograms of total proteins were loaded per lane, and separated by SDS-PAGE and transferred to PVDF membrane. After incubation with secondary antibodies, the expression levels of HIF-1α, S100A1, p-iNOS, iNOS, p-eNOS, eNOS, caspase3,Bax, Bcl-2 and β-actin were visualized using chemiluminescence method. A represent Western blots in panel (1, 2, 3, 4, 5, 6, 7, 8 represent control group, OGD group, SYB group, antimiR-199a group, antimiR-199a+SYB group, L-NAME group, L-NAME+SYB group and antimiR-199a+L-NAME+SYB group respectively). (A1, A2, A3, A4, A5, A6) represent the relative ratio of p-iNOS/iNOS and p-eNOS/eNOS, HIF-1α, S100A1,relative ratio of Bax/Bcl-2, caspase3 respectively. Data were presented as mean ± S.D. (n = 3). One-way ANOVA test was used to determine statistical significance. *P < 0.05 or**P < 0.01 vs. control group, #P < 0.05 or ##P < 0.01 vs. OGD group, ∆P < 0.05 or D∆P < 0.01 vs. SYB+antimiR-199a, +P< 0.05 or ++P < 0.01 vs. L-NAME+SYB.
Fig 3: Effect of safflor yellow B on target protein expression in vascular endotheliumsRats were used to establish hypoxia model. The vascular endotheliums of thoracic aorta were obtained by using cell scraper to gently scratch. Total protein in RAECs from different groups were extracted and measured. Thirty micrograms of total proteins were loaded per lane, and separated by SDS-PAGE and transferred to PVDF membrane. After incubation with secondary antibodies, the expression levels of HIF-1α, S100A1, p-iNOS, iNOS, p-eNOS, eNOS, caspase3, Bax, Bcl-2 and β-actin were visualized using chemiluminescence method. A. represents Western blots in panel (1, 2, 3 represent normal group, hypoxia group, SYB group respectively). A1, A2, A3, A4, A5, and A6 represent the relative ratio of p-iNOS/iNOS and p-eNOS/eNOS, HIF-1α, S100A1,relative ratio of Bax/Bcl-2, caspase3 respectively. Data were presented as mean ± S.D. (n = 3). One-way ANOVA test was used to determine statistical significance. **P < 0.01 vs. normal group, ##P < 0.01 vs. hypoxia group.
Supplier Page from MilliporeSigma for Anti-S100 A1, N-Terminal antibody produced in rabbit