Reperfusion. N, damaging manage stain. Arrows indicate immunopositive cells. Scale bar = 50 m.Figure 5 Effects of EA at acupoints on pp90RSK, active caspase-3-NeuN, and TUNEL expression. (A) Representative photographs showed pp90RSK expression, (B) active caspase-3 (red) colocalizing with NeuN (blue), and (C) TUNEL-positive cells within the ischemic cortex in the sham, model, EA, non-acup, and U0126 + EA groups just after 3 d of reperfusion. N, negative handle stain. Arrows in (A) and (C) indicate pp90RSK- and TUNEL-positive cells, respectively. Arrow in (B) indicates active caspase-3-NeuN double-labeled cells (purple), shown at larger magnification in the bottom suitable panel. Scale bar = 50 m.Cheng et al. BMC Complementary and Alternative Medicine 2014, 14:92 http://biomedcentral/1472-6882/14/Page 8 ofTable 1 Expression of BDNF-, pRaf-1-, pMEK1/2-, pERK1/2-, pp90RSK-, and TUNEL-positive cells (counts/1 mm2)Sham BDNF pRaf-1 pMEK1/2 pERK1/2 pp90RSK TUNEL 0? 0? 0? 0? 0? 0? Model 186 ?46* 149 ?39* 165 ?54* 257 ?52* 261 ?107* 365 ?88* EA 413 ?60*# 348 ?55*# 274 ?31*# 417 ?36*# 497 ?31*# 209 ?59*# Non-acup 178 ?51* 187 ?67* 149 ?52* 219 ?57* 302 ?98* 356 ?80* U0126 + EA 393 ?63*# 338 ?25*# 83 ?54* 198 ?35* 196 ?50* 339 ?79*model group (P 0.05; Figure 5C; Table 1). The amount of TUNEL-positive cells inside the model, non-acup, and U0126 + EA groups showed no important distinction (P 0.05; Figure 5C; Table 1).Effects of EA at acupoints on cytosolic expression of pMEK1/2, pERK1/2, pp90RSK, and pBadMean ?SD (n = 5 or 6). Sham, sham group; Model, model group; EA, EA group; Non-acup, non-acup group; U0126 + EA, U0126 + EA group. *P 0.05 vs. sham group; #P 0.05 vs. model group.Effects of EA at acupoints on the expression of TUNELpositive cellsWe observed increased TUNEL positivity in the ischemic cortex in the model, EA, non-acup, and U0126 + EA group (P 0.05 vs. sham group; Figure 5C; Table 1) after three d of reperfusion. Within the EA group, however, TUNEL positivity was decreased substantially compared with theIn western blot evaluation, we observed that the cytosolic expression of pMEK1/2 within the ischemic cortex immediately after three d of reperfusion showed no substantial difference among the model, non-acup, and U0126 + EA groups (P 0.05). Having said that, cytosolic pMEK1/2 expression was considerably larger inside the EA group than that inside the model group (two.7-fold, P 0.05; Figure 6A and B). We also evaluated the expression of pERK1/2, the downstream target of pMEK1/2, observing substantially greater cytosolic pERK1/2 expression within the EA group compared with all the model group (1.3,3′-Oxybis(propan-1-ol) web 8-fold, P 0.17193-29-2 site 05; Figure 6A and C).PMID:24914310 Cytosolic pERK1/2 expression amongst the model, non-acup, and U0126 + EA groups showed no substantial differenceFigure six Effects of EA at acupoints on cytosolic expression of pMEK1/2, pERK1/2, pp90RSK, and pBad. (A) Representative western blot pictures showed the cytosolic expression of pMEK1/2, pERK1/2, pp90RSK, and pBad in the ischemic cortex in the sham, model, EA, non-acup, and U0126 + EA groups immediately after 3 d of reperfusion. Actin was utilised as an internal handle. The relative cytosolic expression of (B) pMEK1/2, (C) pERK1/2, (D) pp90RSK, and (E) pBad (n = four) was assessed in the ischemic cortex inside the sham, model, EA, non-acup, and U0126 + EA groups. Data are presented as mean ?SD. #P 0.05 compared with the model group.Cheng et al. BMC Complementary and Option Medicine 2014, 14:92 http://biomedcentral/1472-6882/14/Page 9 of(P 0.05). Cytosolic pp90RSK expression was significa.