Ananya Chakravarti, Ph.D.

1) Designed and interpreted translationally relevant animal models of Pulmonary Hypertension and Acute Lung Injury. 2) Delineate the appropriate ranges of experimental parameters to ensure robust, consistent and error-free results. 3) Trained to work in a CL-2 facility. 4) Animal handling including giving intraperitoneal, subcutaneous and intratracheal injections and harvesting organs like heart, lung and spleen for end-point studies. Specific skill sets: #mammalian cell biology #assay development #RT-qPCR #Western Blotting #tissue culture #innovator #team player #conflict resolver #science outreach volunteer

Investigated the genetic and molecular machinery underlying the isoform-specific role of the tumor-suppressor gene, p53 in development using various methods and strategies including but not limited to genetics, cellular biology, and molecular biology. • Developed assays examining gene expression by qRT-PCR, flow cytometry and FACS. • Gene editing using CRISPR/Cas9 for generating and characterizing knockouts of p53 gene. Through collaborative help from other labs, performed functional assays of these mutants leading to a manuscript currently under revision at eLife. • Determined target protein localization (immuno-labeling), ploidy (DNA labeling and flow cytometry), how many cells were actively cycling (EdU incorporation), DNA damage with and without irradiation stress (-H2Av labeling) and cell death (cleaved Caspase labeling and TUNEL), in target cell populations by flow cytometry and confocal microscopy. • Characterized functions and interactions of targeted proteins by overexpression and RNAi, in vivo and in vitro. Downstream analyses of these experiments were done through immunolabeling, SDS-PAGE, and western blotting. Analysis of sequencing was also performed in collaboration. • Peer-reviewed primary literature articles for research journals like Scientific Reports. • Performed general lab management duties like ordering supplies/ maintaining inventories and decision making on effective cost to productivity ratio especially in between research grant renewals. • Honed scientific communication and presentation skills leading to the ‘Women in Science’ fellowship and ‘Honorary Mention’ award in international and regional conferences. • Undertook mentoring and leadership roles including training of four graduate students and one research associate. • Developed active organizational skills leading to smooth operation of the departmental seminars and learned effective time-management which greatly helped in my science outreach activities.

Investigated the role of the evolutionary conserved CCAAT-Binding Factor in the oxidative stress response of the opportunist human pathogen Candida albicans. • Developed fluorescent reporter constructs as well as truncated and fusion protein constructs by molecular cloning approaches and site-directed mutagenesis. • Characterized promoter activity through Renilla luciferase assay. • Characterized protein-protein interactions through yeast-two hybrid system. • Standardized the protocols for various stress assays like catalase enzymatic assay and hydrogen peroxide sensitivity assay to measure the virulence of the different pathogenic strains. • Analyzed gene expression using northern blot and radioactive P-32. • Generated growth curves for uncharacterized mutant strains using spectrophotometer and hemocytometer. • Assisted colleagues in their projects focusing on screening of Histatin-5 derived peptides as potential antifungal drugs against C. albicans using 3T3 fibroblast cell culture system. • Provided intellectual guidance to an undergraduate student and together analyzed and published results in PLoS One.