https://libjncir.jncasr.ac.in/xmlui/handle/10572/1501 2026-04-04T05:31:47Z 2026-04-04T05:31:47Z Khadilkar, Rohan J. Rodrigues, Diana Mote, Ridim Dadasaheb Sinha, Arghyashree Roychowdhury Kulkarni, Vani Magadi, Srivathsa Subramanya Inamdar, Maneesha S. https://libjncir.jncasr.ac.in/xmlui/handle/10572/2477 2017-02-21T10:24:18Z 2014-01-01T00:00:00Z

ARF1-GTP regulates Asrij to provide endocytic control of Drosophila blood cell homeostasis Khadilkar, Rohan J.; Rodrigues, Diana; Mote, Ridim Dadasaheb; Sinha, Arghyashree Roychowdhury; Kulkarni, Vani; Magadi, Srivathsa Subramanya; Inamdar, Maneesha S. Drosophila melanogaster larval hematopoiesis is a well-established model to study mechanisms that regulate hematopoietic niche maintenance and control of blood cell precursor (prohemocyte) differentiation. Molecules that perturb niche function affect the balance between prohemocytes and differentiated hemocytes. The conserved hemocyte-specific endosomal protein Asrij is essential for niche function and prohemocyte maintenance. Elucidating how subcellular trafficking molecules can regulate signaling presents an important challenge. Here we show that Asrij function is mediated by the Ras family GTPase Arf79F, the Drosophila homolog of ADP ribosylation factor 1 (ARF1), essential for clathrin coat assembly, Golgi architecture, and vesicular trafficking. ARF1 is expressed in the larval lymph gland and in circulating hemocytes and interacts with Asrij. ARF1-depleted lymph glands show loss of niche cells and prohemocyte maintenance with increased differentiation. Inhibiting ARF1 activation by knocking down its guanine nucleotide exchange factor (Gartenzwerg) or overexpressing its GTPAse-activating protein showed that ARF1-GTP is essential for regulating niche size and maintaining stemness. Activated ARF1 regulates Asrij levels in blood cells thereby mediating Asrij function. Asrij controls crystal cell differentiation by affecting Notch trafficking. ARF1 perturbation also leads to aberrant Notch trafficking and the Notch intracellular domain is stalled in sorting endosomes. Thus, ARF1 can regulate Drosophila blood cell homeostasis by regulating Asrij endocytic function. ARF1 also regulates signals arising from the niche and differentiated cells by integrating the insulin-mediated and PDGF-VEGF receptor signaling pathways. We propose that the conserved ARF1-Asrij endocytic axis modulates signals that govern hematopoietic development. Thus, Asrij affords tissue-specific control of global mechanisms involved in molecular traffic. Restricted Access

2014-01-01T00:00:00Z Shetty, Deeti K. Inamdar, Maneesha S. https://libjncir.jncasr.ac.in/xmlui/handle/10572/2188 2017-02-21T10:22:43Z 2016-01-01T00:00:00Z

Generation of transgenic human embryonic stem cell line BJNhem20-OCIAD1-OV Shetty, Deeti K.; Inamdar, Maneesha S. Ovarian Carcinoma Immuno-reactive Antigen domain containing protein 1 (OCIAD1) was overexpressed in BJNhem20 human embryonic stem cell line (hESC) using plasmid transfection, followed by stable cell line generation. The construct pCAG-OCIAD1 was introduced into hESCs by microporation. (C) 2015 Published by Elsevier B.V. Restricted Access

2016-01-01T00:00:00Z Shetty, Ronak Inamdar, Maneesha S. https://libjncir.jncasr.ac.in/xmlui/handle/10572/2190 2017-02-21T10:22:46Z 2016-01-01T00:00:00Z

Generation of a constitutively expressing Tetracycline repressor (TetR) human embryonic stem cell line BJNhem20-TetR Shetty, Ronak; Inamdar, Maneesha S. Human embryonic stem cell line BJNhem20-TetR was generated using non-viral method. The construct pCAG-TetRnls was transfected using microporation procedure. BJNhem20-TetR can subsequently be transfected with any vector harbouring a TetO (Tet operator) sequence to generate doxycycline based inducible line. For example, in human embryonic stem cells, the pSuperior based TetO system has been transfected into a TetR containing line to generate OCT4 knockdown cell line (Zafarana et al., 2009). Thus BJNhem20-TetR can be used as a tool to perturb gene expression in human embryonic stem cells. (C) 2016 The Authors. Published by Elsevier B.V. Restricted Access

2016-01-01T00:00:00Z Shetty, Deeti K. Inamdar, Maneesha S. https://libjncir.jncasr.ac.in/xmlui/handle/10572/2186 2017-02-21T10:23:04Z 2016-01-01T00:00:00Z

Generation of a heterozygous knockout human embryonic stem cell line for the OCIAD1 locus using CRISPR/CAS9 mediated targeting: BJNhem20-OCIAD1-CRISPR-39 Shetty, Deeti K.; Inamdar, Maneesha S. Ovarian carcinoma immuno-reactive antigen domain containing 1 (OCIAD1) single copy was knocked out generating an OCIAD1 heterozygous knockout human embryonic stem line named BJNhem20-OCIAD1-CRISPR-39. The line was generated using CRISPR-Cas9D10A double nickase knockout strategy (Mali et al., 2013). (C) 2016 The Authors. Published by Elsevier B.V. Restricted Access

2016-01-01T00:00:00Z