Fig 1: Effects of 1.5% ISPOC, TGF-β1 inhibitor LY2157299, p-Cx43 inhibitor Ro318220, and p-Cx43 activator 18β-GA on neuronal damage in the hippocampal CA1 region after cerebral ischemia/reperfusion (Nissl staining). (a) Brain sections were stained with the Nissl staining solution. The number of Nissl bodies in the hippocampal CA1 area neurons showed the extent of damage. Original magnification: 200x. Scale bars: 50 μm. (b) Number of Nissl bodies in the CA1 area in each group. The results are expressed as means ± SEM (n = 6). ∗P < 0.05, ∗∗P < 0.01.
Fig 2: Effects of 1.5% ISPOC, TGF-β1 inhibitor LY2157299, p-Cx43 inhibitor Ro318220, and p-Cx43 activator 18β-GA on the survival of hippocampal CA1 neurons after cerebral I/R injury in rats (HE staining). (a) Brain sections were stained with HE. Contraction of the nucleus and cellular edema showed the impaired neurons in the hippocampal CA1 area. Original magnification: 200x. Scale bars: 50 μm. (b) Proportion of injured neurons in the CA1 area. The results are expressed as the means ± SEM (n = 8). ∗P < 0.05, ∗∗P < 0.01.
Fig 3: mRNA synthesis level of TGF-ß1 and Cx43 was quantitated by the 2-??Ct method, and ß-actin was used as a reference. The total ribonucleic acid (RNA) was extracted from the hippocampus tissue. (a) Expression of Cx43 mRNA in the ISO group has no significant difference from that in the I/R group. (b) Expression of TGF-ß1 mRNA in the ISO group was higher than that in the I/R group. *P < 0.05, **P < 0.01.
Fig 4: Relationship between p-Cx43 and TGF-β1/Smad2/3 signaling pathway after 1.5% ISPOC in rats with cerebral I/R injury. (a) IF showed fluorescence intensity of p-Cx43 after using the p-Cx43 inhibitor Ro318220 and the blocker of TGF-β1 (original magnification: 200x; ⟶ indicates positions of p-Cx43). (b) Mean fluorescence intensity was used to represent the relative expression level of p-Cx43 after immunofluorescence staining. ∗P < 0.05, ∗∗P < 0.01.
Fig 5: Expression of p-Cx43, Cx43, and TGF-ß1 in the hippocampal tissues when the LY2157299 and 18ß-GA on 1.5% ISPOC were administered to rats with cerebral ischemia/reperfusion injury. (a) Western blot analysis presented the expression of p-Cx43, Cx43, and TGF-ß1 in the hippocampal tissues. ß-Actin was used as an internal control. n = 10 rats per group. (b) The expression pattern of the phosphorylated Cx43 was synchronized with that of TGF-ß1 during the entire process, whereas the total Cx43 protein expression levels did not change in all groups. Dunnett's t-test was used for data analysis. The data are all expressed as the means ± SEM (n = 6). *P < 0.05, **P < 0.01.
Supplier Page from Abcam for Anti-Connexin 43 / GJA1 (phospho S368) antibody