Journal: Acta Pharmaceutica Sinica. B

Article Title: Highly expressed SERCA2 triggers tumor cell autophagy and is a druggable vulnerability in triple-negative breast cancer

doi: 10.1016/j.apsb.2022.05.009

Figure Lengend Snippet: SERCA2 tethers the ER with the isolation membrane through interaction with the ULK1/FIP200 complex and LC3B. (A) Co-IP of FLAG-SERCA2 with the ULK1/FIP200 complex members in transfected SUM1315 cells. (B) Co-localization of DFCP1 with SERCA2 in SUM1315 cells treated with or without EBSS for 1 h. Left panel, Representative images. Scale bar, 5 μm. Right panel, quantification of co-localization. n = 200 puncta. (C) Co-IP of LC3B with the ULK1/FIP200 complex members in SUM1315 cells transiently transfected with the indicated plasmids. (D) Western blot analysis of indicated proteins in SUM1315 cells transfected with the indicated siRNA or plasmids, which were then treated with EBSS for 12 h. (E) Co-localization of FIP200 with LC3 in SUM1315 cells transfected with the indicated siRNA or plasmids, which were then treated with EBSS for 1 h. Upper panel, representative images. Scale bars, 5 μm; insets, 2 μm. Lower panel, quantification of co-localization. n = 200 puncta. (F) Co-localization of DFCP1 with LC3 in SUM1315 cells transfected with the indicated plasmids, which were then treated with EBSS for 4 h in the absence or presence of 100 nmol/L thapsigargin (TG). Upper panel, representative images. Scale bars, 5 μm; insets, 2 μm. Lower panel, quantification of co-localization. n = 200 puncta. Data are shown as the mean ± SD of at least three independent experiments. ∗ P < 0.05, ∗∗∗ P < 0.001.

Article Snippet: Anti-LC3B (3868), anti-SERCA2 (9580), anti-WIPI2 (8567), anti-calnexin (2679) antibodies and Autophagy Induction (ULK1 Complex) Antibody Sampler Kit (46486) were purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Isolation, Co-Immunoprecipitation Assay, Transfection, Western Blot

Journal: Frontiers in Cellular Neuroscience

Article Title: Electroacupuncture Inhibits Neuronal Autophagy and Apoptosis via the PI3K/AKT Pathway Following Ischemic Stroke

doi: 10.3389/fncel.2020.00134

Figure Lengend Snippet: Effect of EA intervention on the protein of the PI3K/AKT pathway in cerebral I/R-injured rats. (A) Western blot analysis showing the levels of PI3K, Atg7, P-AKT, t-AKT, P-mTOR, and t-mTOR among the sham, MCAO/R, EA, EA+NC, and EA+D groups. (B) Statistical analysis showing the difference of PI3K in the ipsilateral hippocampus of the rats ( n = 5/group). (C) Statistical analysis showing the expression of Atg7 in the ipsilateral hippocampus of the rats. (D) Statistical analysis showing the level of P-mTOR/t-mTOR in the ipsilateral hippocampus of the rats. (E) Statistical analysis showing the level of P-AKT/t-AKT in the ipsilateral hippocampus of the rats.

Article Snippet: After blocking in a 5% bovine serum albumin solution for 1 h at 37°C, the membranes were incubated at 4°C overnight with primary antibodies against LC3 (1:500, #18725-1-AP, proteintech), P62 (1:1,000, #18420-1-AP, proteintech), LAMP-1 (1:1,000, #62562, Abcam), cleaved-caspase3 (1:500, #66470-2-lg, proteintech), Atg7 (1:1,000, #32345, Cell Signalling Technology, Danvers, MA, USA), PI3K (1:1,000, #4249, Cell Signalling Technology, Danvers, MA, USA), mTOR (1:1,000, #2972 Cell Signalling Technology, Danvers, MA, USA), Phospho-mTOR (1:1,000, #5536, Cell Signalling Technology, Danvers, MA, USA), AKT (1:1,000, #4691, Cell Signalling Technology, Danvers, MA, USA), Phospho-AKT (Thr308, #13038, Cell Signalling Technology, Danvers, MA, USA) and GAPDH (1:5,000, #10494-1-AP, proteintech).

Techniques: Western Blot, Expressing