Journal: Transgenic research

Article Title: High efficiency plastid transformation in potato and regulation of transgene expression in leaves and tubers by alternative 5' and 3' regulatory sequences.

doi: 10.1007/s11248-010-9402-9

Figure Lengend Snippet: Fig. 3 Expression of gfp in tubers of transplastomic potato lines, obtained with ten different transformation constructs as indicated, and non-transformed De´sire´e plants (WT). a Northern blot analysis of total RNA (20 lg) probed with [a- 32P]dCTP labelled gfp probe. Hybridizations with 18S radiolabelled probe was used as loading control. b Western blot analysis of total soluble proteins (50 lg) using anti-GFP antibodies. The recombinant GFP protein in 50 lg of WT protein from tubers was used as standard

Article Snippet: Different amounts (50, 25, 10 and 2.5 ng) of purified recombinant GFP protein (rGFP, Clontech, Mountain View, CA, USA) were added to 20 lg protein from leaves or 50 lg protein from tubers of non-transformed Désirée plants and used as a standards.

Techniques: Expressing, Transformation Assay, Construct, Northern Blot, Control, Western Blot, Recombinant

Journal: Transgenic research

Article Title: High efficiency plastid transformation in potato and regulation of transgene expression in leaves and tubers by alternative 5' and 3' regulatory sequences.

doi: 10.1007/s11248-010-9402-9

Figure Lengend Snippet: Fig. 2 Expression of gfp in leaves of transplastomic potato lines, obtained with eleven different transformation constructs as indicated, and non-transformed De´sire´e plants (WT). a Northern blot analysis of total RNA (5 lg) probed with [a- 32P]dCTP labelled gfp probe. Hybridizations with psbD radiolabelled probe was used as loading control. b Western blot analysis of total soluble proteins (10 lg for pVL12, pVL13 and pVL15 vectors, and 20 lg for all other constructs) using anti-GFP antibodies. The recombinant GFP protein in 20 lg of WT protein from leaves was used as standard

Article Snippet: Different amounts (50, 25, 10 and 2.5 ng) of purified recombinant GFP protein (rGFP, Clontech, Mountain View, CA, USA) were added to 20 lg protein from leaves or 50 lg protein from tubers of non-transformed Désirée plants and used as a standards.

Techniques: Expressing, Transformation Assay, Construct, Northern Blot, Control, Western Blot, Recombinant

Journal: Leukemia

Article Title: GNAS knockout potentiates HDAC3 inhibition through viral mimicry-related interferon responses in lymphoma.

doi: 10.1038/s41375-024-02325-4

Figure Lengend Snippet: Fig. 4 GNAS KO synergizes with HDAC3 inhibition to enhance CD8+ T cell-induced cytotoxicity. A Schematic representation of the T cell co-culture cytotoxicity assay. B Quantification of percentage cell death in DLBCL cells. Error bars represent ± SEM (n = 2 independent experiments). *, p < 0.05; **, p < 0.01; ***, p < 0.001 (log2 transformed values or values were compared; one-way ANOVA with Tukey’s HSD test for multiple comparisons). C Flow cytometry-based determination of cell death. Single cells and non-debris were included for analysis of 5- (and 6)-Carboxyfluorescein diacetate succinimidyl ester (CFSE) and propidium iodide (PI) signals. CFSE positive and negative populations represent DLBCL cells and CD8+ T cells, respectively. PI positive and negative groups represent dead and live cells, respectively. The total number of single DLBCL cells acquired per sample was 10,000. The percentages shown are percentage cell death within the total DLBCL cell population per sample. PBMCs, peripheral blood mononuclear cells.

Article Snippet: The selective HDAC3 inhibitors RGFP966 and BRD3308 (Selleckchem), the pan-HDAC inhibitor romidepsin (Selleckchem), the cAMP analog 8-Br-cAMP (Tocris), the TBK1 degrader TBK1 PROTAC 3i (Tocris), the JAK1/2 inhibitor ruxolitinib (Selleckchem), and the DNA methyltransferase inhibitor 5-Aza2’-deoxycytidine (5-AZA-CdR; MedChemExpress) were reconstituted and stored according to the manufacturer’s instructions.

Techniques: Inhibition, Co-Culture Assay, Cytotoxicity Assay, Transformation Assay, Flow Cytometry

Journal: Leukemia

Article Title: GNAS knockout potentiates HDAC3 inhibition through viral mimicry-related interferon responses in lymphoma.

doi: 10.1038/s41375-024-02325-4

Figure Lengend Snippet: Fig. 5 GNAS KO plus RGFP966 is associated with dsRNA formation and TE expression for viral mimicry induction. A Representative images of SU-DHL-4 cells stained by Hoechst 33342 (for cell nuclei; blue) and J2 antibody (for dsRNAs; green) from immunofluorescence analysis. Control LacZ cells omitting primary J2 antibody and treated with 300 nM 5-AZA-CdR were the negative and positive control, respectively. Scale bar, 25 µm. B Quantification of results in A. **p < 0.01; ****p < 0.0001 (Kruskal–Wallis test with Wilcoxon rank sum test for pairwise comparisons). C Volcano plots showing normalized expression dynamics of differentially expressed TEs (FDR < 0.05 and |Log2 fold change | > 0) identified from TEtranscripts analysis of cell-line bulk RNA-seq data. Upregulated and downregulated TEs are highlighted in red and blue, respectively. The horizontal and vertical dotted lines indicate FDR = 0.05 and Log2 fold change = 0, respectively. D Normalized TE expression under each comparison by a ratio of upregulated versus downregulated TEs shown in C. The dotted line represents normalized TE expression = 1 (equal numbers of upregulated and downregulated TEs).

Article Snippet: The selective HDAC3 inhibitors RGFP966 and BRD3308 (Selleckchem), the pan-HDAC inhibitor romidepsin (Selleckchem), the cAMP analog 8-Br-cAMP (Tocris), the TBK1 degrader TBK1 PROTAC 3i (Tocris), the JAK1/2 inhibitor ruxolitinib (Selleckchem), and the DNA methyltransferase inhibitor 5-Aza2’-deoxycytidine (5-AZA-CdR; MedChemExpress) were reconstituted and stored according to the manufacturer’s instructions.

Techniques: Expressing, Staining, Control, Positive Control, RNA Sequencing, Comparison