Journal: Science Advances

Article Title: Artificial cells drive neural differentiation

doi: 10.1126/sciadv.abb4920

Figure Lengend Snippet: The synthesis of PFO monomers (light orange) is controlled by a genetic AND-gate that requires both LuxR and 3OC6 HSL for gene expression. Monomers of PFO assemble into pores in the presence of cholesterol-containing membranes, thereby releasing BDNF. Homodimers of mature BDNF (green) act on the cognate receptor TrkB, activating signaling pathways leading to neural stem cell differentiation and maturation. The figure was not drawn to scale.

Article Snippet: Human TrkB complementary DNA (cDNA) was synthesized by GenScript in two parts, joined in-house by Gibson Assembly, and cloned into a pLenti vector with standard molecular cloning techniques.

Techniques: Gene Expression, Protein-Protein interactions, Cell Differentiation

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Novel function of N -acetyltransferase for microtubule stability and JNK signaling in Drosophila organ development

doi: 10.1073/pnas.2010140118

Figure Lengend Snippet: Acetylation activity of Mnat9 is not required for its function in JNK signaling and microtubule stability. (A) Mnat9 has the acetyl-CoA binding motif conserved in all known NATs. (B) Microtubule turbidity assay results show that the addition of either Mnat9 or Mnat9[AcDel] increases the speed of microtubule polymerization. (C–F) Effects of Mna9 overexpression in the notum. Overexpression of wild-type Mnat9 shows a weak thorax cleft (D). Overexpression of mutated Mnat9[AAA] (E) or Mnat9[AcDel] (F) shows a similar thorax cleft phenotype as the wild type. (G–I) Rescue of Mnat9 RNAi phenotype by C96-Gal4. The wing-notching phenotype caused by Mnat9 RNAi (G) is fully suppressed by overexpressing hNAT9 (H, n = 88) or hNAT9[AAA] (I, n = 44). (J–L) Rescue of Mnat9 RNAi phenotype by en-Gal4. The small and wrinkled wing phenotype caused by Mnat9 RNAi (J) is almost fully suppressed by overexpressing hNAT9 (K, n = 60) or hNAT9[AAA] (L, n = 36). (M, N) Effects of Mnat9 overexpression by en-Gal4 in microtubule stability. Treatment of the wing imaginal disc with demecolcine (50 μM) destabilizes the tubulin pattern. The posterior compartment with overexpression of Mnat9 (M) or Mnat9[AcDel] (N) is more resistant to demecolcine than the anterior control region. [Scale bars: 200 μm (C–F), 0.5 mm (G–L), and 50 μm (M, N).]

Article Snippet: We generated UAS-Mnat9[AAA] and UAS-Mnat9[AcDel] transgenic flies using oligo synthesis service (Twist) and UAS-hNAT9 and UAS-hNAT9[AAA] transgenic flies using human hNAT9 complementary DNA (cDNA) (Genscript).

Techniques: Activity Assay, Binding Assay, Over Expression, Control

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Novel function of N -acetyltransferase for microtubule stability and JNK signaling in Drosophila organ development

doi: 10.1073/pnas.2010140118

Figure Lengend Snippet: Overexpression of Mnat9 suppresses JNK activity. puc-lacZ expression in the notum region of the wing disc (A) was strongly suppressed by overexpressing Mnat9 (B, white arrow). The overexpressed dominant-negative form of Kay protein causes the notched wing phenotype in adult male wings (D). Ectopic expression of Mnat9 enhances the phenotype (E, E′), while a single expression of Mnat9 shows normal adult wings (C). Overexpression of Mnat9 or human homolog hNAT9 causes a mild thorax closure defect phenotype (F, F′). Mutant or down-regulation of JNK signaling components further enhances the Mnat9 overexpression phenotype (G–J′). [Scale bars: 50 μm (A, B), 0.5 mm (C–E′), and 200 μm (F–J′).]

Article Snippet: We generated UAS-Mnat9[AAA] and UAS-Mnat9[AcDel] transgenic flies using oligo synthesis service (Twist) and UAS-hNAT9 and UAS-hNAT9[AAA] transgenic flies using human hNAT9 complementary DNA (cDNA) (Genscript).

Techniques: Over Expression, Activity Assay, Expressing, Dominant Negative Mutation, Mutagenesis