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. 2020 May 22;10:8587. doi: 10.1038/s41598-020-65468-1

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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I. Ultrasound exposure over the right tumor spatially enhanced the uptake of the NERh6g and CLSM image of a thin section of the tumors. Red colour fluorescence is due to the internalization of NERh6g by tumor, (A) central region of the tumor (B) peripheral region of the tumor (C) peripheral region of the left tumor which was not exposed to ultrasound (D) peripheral region of the right tumor which was exposed to ultrasound for 60 sec at intensity 2 W/cm², duty cycle of 50%, (E) central region of the left tumor, (F) central region of the right tumor. II. The average pixel intensity of the red fluorescence from each image was calculated and plotted. Ten points were considered from each image for calculating pixel intensity. P value was calculated by Student t-test, and values are presented in the form of mean ± SD. ****P ≤ 0.0001 and **P ≤ 0.01. III. Antitumor efficacy of the Cur_NE in the presence and absence of MB injection. The right tumor was exposed to ultrasound whereas the left tumor was kept unexposed. (A) A graph is showing change in tumor volume over time after oral administration of formulations. (B,C) Relative tumor growth was observed over the period. (D) Image of tumors from different groups at the end of treatment (30^th day), (i) PBS (ii) Cur free (iii) Cur_NE (iv) Cur_NE + MB. (E) Kaplan-Meier curve, percent survival of mice in different groups of treatment (Log-Rank test trend *P ≤ 0.04). (F) Change in average body weight in each group over the entire period of study. P value was calculated by Student t- test, and data are presented as mean ± SD.