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Image Search Results
Journal: Nature Communications
Article Title: Targeting miR-126 in inv(16) acute myeloid leukemia inhibits leukemia development and leukemia stem cell maintenance
doi: 10.1038/s41467-021-26420-7
Figure Lengend Snippet: a Western blot analysis of HDAC8 in 32D-CM cells with NS or Hdac8 shRNA (#1, #2) (left). Relative expression of pre-miR-126 and mature miR-126 in 32D-CM cells with NS (red) vs. Hdac8 shRNA#1 (light blue), Hdac8 shRNA#2 (dark blue) (right). b Relative expression in 32D-CM cells treated with vehicle (red) vs. HDAC8i (22d 2.5 μM or 5 μM; purple) for pre-miR-126 (left; 2.5 μM p < 0.0001; 5 μM p < 0.0001) and miR-126 (right; 2.5 μM p = 0.012; 5 μM p = 0.0055). c Representative image of IF co-staining of RAN (green) and HDAC8 (red) in 32D cells (left; scale bar 10 μm). IP with anti-IgG control or anti-HDAC8 followed by immunoblotting (IB) with anti-RAN antibodies (right). Representative results of two independent experiments with similar results are shown. d IP with anti-RAN and IB with antibodies for Ac-Lys, RCC1, XPO5, or RAN in 32D-CM cells not-treated (NT) or treated with DMSO vehicle or 22d (2 µM) (left). The levels of RAN-GTP determined by RAN activation assay (right). The densitometry of RAN-GTP levels measured from three assays are shown on the bottom. e IP with anti-RAN and IB with antibodies for Ac-Lys, RCC1, XPO5, or RAN (left), and RAN activation assay (right) in 32D-CM cells not-treated (NT) or transduced with shCtrl control or shCM for 24 h. f IP with anti-RAN and IB with antibodies for Ac-Lys, RCC1, XPO5, or RAN (left), and RAN activation assay (right) of BM cells isolated from Ctrl or CM mice. g Schematic model of the mechanism by which CM regulates miR-126/EGFL7 transcription as well as miR-126 biogenesis through enhancing HDAC8 activity which in turn promotes RAN-XPO5 mediated transportation. Pre-miR-126 was normalized to B2m and mature miR-126 was normalized to snoRNA234 . Data are presented as the mean ± SD and statistical significance shown was determined using two-tailed Student’s T tests (* p < 0.05; ** p < 0.01; *** p < 0.001).
Article Snippet:
Techniques: Western Blot, shRNA, Expressing, Staining, Control, Activation Assay, Transduction, Isolation, Activity Assay, Two Tailed Test