Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)
Article Title: Nuclear Control of Mitochondrial Homeostasis and Venetoclax Efficacy in AML via COX4I1.
doi: 10.1002/advs.202404620
Figure Lengend Snippet: Figure 5. COX4I1 governs pivotal stages of Complex IV assembly. A) Schematic depiction of the COX4I1 high-resolution CRISPR tiling library screen conducted in Molm13-Cas9+ cells. B) 2D annotation illustrating COX4I1 peptide position (x-axis) and CRISPR tiling score (y-axis). The red line represents the smoothed model of the CRISPR scan score derived from individual sgRNAs (dots) targeting the coding exons of COX4I1. The CRISPR hypersensitive P1 and P2 regions are indicated. C,D) Impact of wild-type (WT), ΔP1, and ΔP2 COX4I1 expression on (C) cell number expansion and (D) mitochon- drial respiration in COX4I1-KO Molm13 cells (n = 3 for each group). E) Western blot analysis of COX4I1, COX5A, MT-CO2, COX6B1, and 𝛽-actin in
Article Snippet: PVDF membranes were subsequently blocked with 5% bovine serum albumin (Fisher Scientific) in Tris-buffer saline-tween 20 (TBST) at room temperature for 1 h. Primary antibodies targeting COX4I1 (rabbit mAb [clone 3E11]; 4850, Cell Signaling Technology, 1:1000), OPA1 (rabbit mAb [clone D6U6N]; 80471, Cell Signaling Technology, 1:1000), cytochrome c (rabbit mAb [clone EPR1327]; ab133504, Abcam, 1:1000), VDAC (rabbit mAb [clone D73D12]; 4661, Cell Signaling Technology, 1:1000), COX5A (rabbit mAb [clone EPR14208(B)]; ab180129, Abcam, 1:1000), MT-CO2 (rabbit mAb [clone E6U9K]; 50003, Cell Signaling Technology, 1:1000), COX6B1 (mouse mAb [clone 3F9D3D11AF6]; ab110266, Abcam, 1:500), cleaved caspase 3 (rabbit mAb [clone 5A1E]; 9664, Cell Signaling Technology, 1:1000), BAX (rabbit mAb [clone D2E11], 5023, Cell Signaling Technology, 1:1000), BAK (rabbit mAb [clone D4E4], 12105, Cell Signaling Technology, 1:1000), and β-actin (mouse mAb [clone 8H10D10]; 3700, Cell Signaling Technology; 1:1000) were then applied to the membranes at 4 °C overnight.
Techniques: CRISPR, Derivative Assay, Expressing, Western Blot
