Fig 2: Immunostaining for the expression of Cx43, HC1, and Casp3 in rat’s brain after HI insult. Cx43, HC1, and Casp3 were stained green, and nuclei were stained blue. Bar = 25 µm. a1-3, The expression of Cx43, HC1, and Casp3 in SVZ from rat without the insult of HI (Cont.); b1-3, The expression of Cx43, HC1, and Casp3 in SVZ from rat 1 hour after HI (T1 hour); c1-3, The expression of Cx43, HC1, and Casp3 in SVZ from rat 12 hours after HI (T12 hour); d1-3, The expression of Cx43, HC1, and Casp3 from rat 24 hours after HI (T24 hour); e1-3, The expression of Cx43, HC1, and Casp3 in SVZ from rat 48 hours after HI (T48 hour); f1, The plaque area (pixels) of Cx43 and HC1 staining per cell with the time points. f2, The percentage of Casp3 positive cells (%) with the time points. *P < .05. Cx43 indicates connexin 43; Casp3, caspase 3; HCl, hemichannel; HI, hypoxia–ischemia; SVZ, subventricular zone.

Fig 3: Western blot analysis of Cx43, HC1, and Casp3 expression from astrocytes following OGD treatment. Astrocytes were treated with OGD techniques. The lysates were resolved by SDS-PAGE, and immunopositive bands were semiquantified by scanning laser densitometry. Data are expressed as mean ± SD corresponding to data displayed, * P < .05. A1-2, Astrocytes cell line; B1-2, Psup astrocytes; C1-2, shRNA astrocytes; D1, Tublin was detected as a control. Cx43 indicates connexin 43; Casp3, caspase 3; HCl, hemichannel; OGD, oxygen–glucose deprivation; shRNA, small hairpin RNA; SDS-PAGE, sodium dodecyl sulfate–polyacrylamide gel electrophoresis; SD, standard deviation.

Fig 4: Caspase-3% on the lymphoreticular organs. Domestic chickens (AC-A) and Alabio ducks (AC-I) infected with ducks/Aceh Besar_IND/2013/eoAC080721 isolate. T: thymus, BF: Bursa of fabricious, L: Spleen, ST: Ceca tonsils.

Fig 5: Caspase-3 expression on the lymphoreticular organs. Domestic chickens (AC-A) and Alabio ducks (AC-I) infected with ducks/Aceh Besar_IND/2013/eoAC080721 isolate. T: thymus, BF: Bursa of fabricious, L: Spleen, K: Cortex, M: Medullar, (*): Plica epithelia, FL: Lymphoid follicle, GM: Germinal center. Immunohistochemical using anti-casp3 antibody.