Abstract
Here, we developed a cell defined siRNA microarray (CDSM) for human bone marrow stromal cells (hBMSCs) designed to control the culture of cells inside the spot area without reducing the efficiency of siRNA silencing, “Development of a cell-defined siRNA microarray for analysis of gene functionin human bone marrow stromal cells” (Kim et al., 2016 [1]). First, we confirmed that p65 protein inhibition efficiency was maintained when hBMSCs were culture for 7 days on the siRNA spot, and siRNA spot activity remained in spite of long term storage (10 days and 2 months). Additionally, we confirmed p65 protein inhibition in U2OS cells after 48 h reverse transfection.
Highlights
Subject area More specific subject area Type of data How data was acquired Data format Experimental factors Experimental features
U2OS cells were carried out by siRNA reverse transfection for 2 days following with same process as human bone marrow stromal cells (hBMSCs) Institut Pasteur Korea, IP-Korea, Seongnam-si, Republic of Korea
This data proves that cell defined siRNA microarray (CDSM) technology can be applied to hBMSCs and U2OS cells to identify gene functions
Summary
Analyzed hBMSCs and U2OS cells were cultured on the array and induced knockdown of p65 by siRNA spot hBMSCs were seeded on siRNA spot array and incubated for 10 min for cell attachment and unattached cells were removed. Cell-spots were incubated for 2 days and 7 days to induce knock down by siRNA spot This experiment was performed for confirmation of siRNA spot activity by reverse transfection time and by storage period of siRNA spot array. U2OS cells were carried out by siRNA reverse transfection for 2 days following with same process as hBMSCs Institut Pasteur Korea, IP-Korea, Seongnam-si, Republic of Korea. This data shows that CDSM is a useful tool for image-based siRNA screening with cellular models. This data shows that the preservation of siRNA stability in CDSM offers opportunity for siRNA screening This data proves that CDSM technology can be applied to hBMSCs and U2OS cells to identify gene functions. U2OS osteosarcoma cells, (ATCC, Manassas, VA, USA) were cultured in DMEM high-glucose medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin in a humidified atmosphere of 5% CO2at 37 °C
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