mcui4  (Tocris)

Journal: Drug Design, Development and Therapy

Article Title: Danggui-Shaoyao-San Can Ameliorate Alzheimer’s Disease by Inhibiting Hippocampal Neuron Apoptosis: Findings from Serum Pharmacology

doi: 10.2147/DDDT.S490900

Figure Lengend Snippet: Effect of DSS serum on apoptosis of HT-22. Cells were divided into five groups: HT-22 (H), HT-22+STZ (S), HT-22+STZ+DSS (D),HT-22+STZ +MCUi4(M), HT-22+STZ+DSS+MCUi4 (D+M). ( A ) Results of Western blot. ( B ) Protein expression results of Bax/Bcl-2(n = 3). ( C ) Results of Western blot. ( D ) Protein expression results of p-CaMKII(n = 3). ( E ) Protein expression results of cyt-c(n = 3). ( F ) Protein expression results of cl-Caspase-3 (n = 3). ( G ) Results of Western blot. ( H )Protein expression results of MCU (n = 3). ( I )Apoptosis by flow cytometry. ( J )Apoptosis rate(n = 3). *indicates P < 0.05; **indicates P < 0.01; *** indicates P < 0.001; **** indicates P < 0.0001; ns indicates no statistical difference.

Article Snippet: The MCU is blocked by treating HT-22 cells with 10μM MCUi4(HY-138620, MCE).

Techniques: Western Blot, Expressing, Flow Cytometry

Journal: Cells

Article Title: Mitochondria-Endoplasmic Reticulum Interplay Regulates Exo-Cytosis in Human Neuroblastoma Cells

doi: 10.3390/cells11030514

Figure Lengend Snippet: Inhibition of ER to mitochondria Ca 2+ shuttling abrogates Mfn2-dependent down-regulation of synaptophysin levels and increased exocytosis ( A ) Schematic representation showing mode of action of Xestospongin C (XeC) and MCUi4 at MERCS ( B ) Representative immunoblots of NC siRNA or Mfn2 siRNA SH-SY5Y cells treated or 6 h with DMSO or 1 µM XeC. Blots were probed with antibodies against Mfn2, synaptophysin and actin, which was used as a loading control. ( C ) Bar graph shows the amounts of synaptophysin protein once standardized to actin content in each sample ( n = 5 independent cultures) ( D ) Representative confocal images of Mfn2 siRNA SH-SY5Y cells treated with DMSO or XeC 1 µM for 6 h. Panels show initial fluorescence (F 0 ) and final fluorescence (F f ). Scale bar = 10 µm ( E ) Graph shows normalized SypHy fluorescence over time in Mfn2 siRNA treated with DMSO or XeC, each timepoint ( F ) was normalized to average pre stimulation fluorescence levels (F pre ). After one minute of basal recording, cells were depolarized with 50 mM KCl ( n = 24–27 cells from four independent cultures). Ratiometric difference between F f /F 0 was shown to be significant p ≤ 0.01. ( F ) Representative immunoblots of NC siRNA or Mfn2 siRNA SH-SY5Y cells treated for 6h with DMSO or 1 µM MCUi4. Blots were probed with antibodies against Mfn2, synaptophysin and actin which was used as a loading control. ( G ) Bar graph shows the amounts of synaptophysin protein once standardized to actin content in each sample ( n = 5 independent cultures). ( H ) Representative confocal images of Mfn2 siRNA SH-SY5Y cells treated with DMSO or MCUi41 µM for 6 h. Panels show initial fluorescence (F 0 ) and final fluorescence (F f ). Scale bar = 10 µm I) Graph shows normalized SypHy fluorescence over time in NC and Mfn2 siRNA treated with DMSO or MCUi4, each timepoint ( F ) was normalized to average pre stimulation fluorescence levels (F pre ). After one minute of basal recording, cells were depolarized with 50 mM KCl ( n = 16–19 cells from 4 independent cultures). Ratiometric difference between F f /F 0 was shown to be significant p ≤ 0.05 between Mfn2 DMSO and Mfn2 MCUi4. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001. Data shown as mean ± SEM.

Article Snippet: Treatment with Xestospongin C (XeC; #1280 Tocris, Bristol, UK) or MCUi4 (#7195, Tocris) 1μM was carried out for 6h before imaging or cell lysis was performed, and control conditions were incubated with an equal volume of DMSO.

Techniques: Inhibition, Western Blot, Control, Fluorescence