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. 2017 Sep 4;131(18):2363–2375. doi: 10.1042/CS20171182

Figure 2. TMZ directly protects RGCs against apoptosis induced by OGD/R.

Figure 2

(A) Immunostaining results indicated that primary RGCs indeed expressed characteristic markers of RGCs including Brn-3a (red), RBPMS (green, left), and β3-tubulin (green, right). (B) CCK8 assay showed that cellular metabolism was affected when TMZ concentration reached 50 µM. (C) CCK8 assay showed that cell survival was significantly increased by TMZ treatment after OGD/R. (D,E) The results of flow cytometry showed that the proportion of apoptotic cells was increased to 15.8% (±3.1%) after OGD/R exposure. TMZ significantly decreased the proportion of PI + Annexin + cells to an average of 2.93%. (F) Western blot analysis demonstrated that TMZ significantly suppressed the level of cleaved caspase-8. (G) The reduction in caspase-8 activity was confirmed by caspase-8 activity assay. Scale bar =20 μm. The data represent the means ± S.D. (n=8); *P<0.05, **P<0.01.