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IRE-1 (ERN1) target sequences are CUACCCAAACAUCGGGAAA, CUCCAGAGAUGCUGAGCGA, AUAAUGAAGGCCUGACGAA, GUCCAGCUGUUGCGAGAAU. Nontargeting control sequences were not provided. JNK1/2 and PERK siRNA (1, # 6232, 2, # 6233) was purchased from Cell Signaling Technology, Inc. (Danvers, MA) and sequences were not provided. Cells at 50 to 60% confluence were Liothyronine Sodium transfect with siRNA (20�C50?nM) using the RNAifect Transfection Regaent (QIAGEN) according to the manufacturer's protocol. Cells were cultured for 48?h, and then treated with vehicle for an additional 48?h. Proteins were then isolated for western blotting. 2.8. Statistical Analysis Statistical analysis was performed by one-way ANOVA. The intergroup comparisons (post-hoc analysis) among the data with equal variances were performed with the least significant difference (LSD) method, while Tamhane's T2 method was used for the data with unequal variances. P selleck chemical cases of SLE patients and 10 cases of healthy donors. We sought to assess apoptotic cells using FCM analysis by Annexin V-FITC/PI staining at the fourth passage. A significant increase was discovered in Annexin V-positive cells among BM-MSCs from SLE patients (32.3 �� 12.0%) compared to normal controls (4.1 �� 3.7%) (P see more of caspase-3 was enhanced in BM-MSCs from SLE patients (Figure 1(c)). All these results showed that BM-MSCs were apoptotic in SLE patients group. Figure 1 Apoptosis of BMSCs from SLE patients. (a) BM-MSCs at passage 4 were isolated and harvested from SLE patients and normal controls, and analyzed by Annexin V-FITC/PI staining. Annexin V-positive cells were 4.1 �� 3.7% in normal controls, 32.3 �� ... 3.2. ERS Was Involved with BM-MSCs from SLE Patients Electron microscopy was used to evaluate the ultrastructural changes of ER in the BM-MSCs. Cells in the control appeared to be normal with relatively healthy-looking ER, mitochondria and nuclei. Dilated and distorted ER, swollen mitochondria, and condensation of chromatin were found in the BM-MSCs from SLE patients; more protein aggregates were seen within the ER lumen than that observed in control groups, indicating severe ERS in BM-MSCs from SLE patients.