Journal: The Journal of Biological Chemistry

Article Title: Dicer represses the interferon response and the double-stranded RNA-activated protein kinase pathway in mouse embryonic stem cells

doi: 10.1016/j.jbc.2021.100264

Figure Lengend Snippet: Effects of PKR inhibition on cell proliferation and cell cycle of ESCs and D−/−ESCs. A , cells were seeded at about 30% confluence and cultured in the absence (Con) or presence of PKR inhibitor C16 at the indicated concentrations. The cell numbers were determined by cell viability assay after 60 h incubation. The cell number in the control (Con) was set as 100%. B , cells were treated under the conditions as described in A for 48 h. Cell cycle was analyzed by flow cytometry. Insets show percentages of cell populations in different phases. Arrows denote the changes of G1 phase cells in control and C16 (1 μM)-treated cells. The histograms are representatives of flow profiles from experiments that were repeated three times with similar results. C , cells were transfected with siRNA against PKR (siPKR) or control siRNA (siCon). After 48 h incubation, the cell number was determined by the method described in A . The number of cells without transfection (Con) was set as 100%. PKR knockdown is assessed by Western blot analysis. The blot inset shows two sets of independent samples. In bar graphs, the values are as mean ± SD of a representative experiment performed in biological triplicate that was performed at least twice ( A ) or the mean ± SD of two combined independent experiments each performed in biological triplicate ( C ). p < 0.0001,∗∗∗∗; p < 0.001,∗∗∗; p < 0.01,∗∗; p < 0.05.∗ Compared groups are indicated by a horizontal bar.

Article Snippet: siRNA targeting PKR and negative control siRNA (Santa Cruz Biotechnology) were transfected to the cells with Endofectin Max at a final concentration of 100 nM.

Techniques: Inhibition, Cell Culture, Viability Assay, Incubation, Control, Flow Cytometry, Transfection, Knockdown, Western Blot