DDIT3/CHOP Antibody - #AF5280

FIGURE 6 PPARγ reduces tunicamycin-induced ER stress by regulating NGBR. (A–C) HUVEC cells in a six-well plate were transfected with control siRNA (NC siRNA) or NGBR siRNA for 24 h in serum-free medium, followed by switching the cells into complete medium to culture for another 24 h. After treatment with rosiglitazone (10 μM) for 12 h, the cells were treated with tunicamycin (0.5 μg/ml) with or without rosiglitazone for another 12 h. Expression of CHOP, BIP, NGBR p-PERK, p-IRE1α and c-ATF6 protein was determined by Western blot (A, B). Expression of CHOP, BIP and NGBR mRNA was determined by qPCR (C). Values were expressed as means ± SD. *p < 0.05; **p < 0.01; ns, not significant (n = 3). (D) liver total proteins were collected from Figure 4. Expression of BIP and CHOP in mouse liver was determined by Western blot. Values were expressed as means ± SD. *p < 0.05; **p < 0.01; ns, not significant (n = 3).

FIGURE 4 CTRP1 protected against cerebral ischemia reperfusion injury via alleviating neuron injury and apoptosis. (A) Neuron injury was analyzed by double labeling immunofluorescence staining. n = 3 per group. The representative images were acquired under × 400 magnification, scale bars = 50 μm. (B) Apoptosis in the cortex was analyzed by TUNEL, n = 3 per group. The representative images were acquired under × 200 magnification, scale bars = 100 μm. (C) Western blot analyzed the expression of CTRP1, BAX, Bcl-2, and CHOP in cortex. n = 4 per group. ****p < 0.0001, ***p < 0.01, **p < 0.01, *p < 0.05 vs. sham group; ####p < 0.0001, ##p < 0.01, #p < 0.05 vs. MCAO/R + LV-NC group.

Fig. 5. Iron overload activated ER stress-mediated apoptosis pathway. (a) and (b) protein expression levels of Bip, ATF4 and CHOP and the phosphorylation level of eIF2α in MC3T3-E1 osteoblast cells treated by indicated concentrations of FAC for 48 h. The data were presented as mean ± SEM, n = 3. *p < 0.05, **p < 0.01, ***p < 0.001 vs. control group. (c) Immunofluorescence analysis of CRT in MC3T3-E1 osteoblast cells using the anti-calreticulin antibody. Cytolemma were counterstained with Dil and nuclei were counterstained with DAPI. (d) Protein expression levels of CRT in MC3T3-E1 osteoblast cells treated by indicated concentrations of FAC for 48 h. The data were presented as mean ± SEM, n = 3. **p < 0.05, **p < 0.01, ***p < 0.001vs. control group.