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. 2021 Jul 16;12:4357. doi: 10.1038/s41467-021-24574-y

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© The Author(s) 2021

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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Fig. 7 — a U251 with inducible US28 expression (No Nb) were transduced with lentiviral vector expressing inducible non-US28 targeting intrabody-mVenus (Irr Nb mV) or VUN103-mVenus (VUN103 mV). Cells were (induced) or were not (Non-induced) induced with doxycycline. US28 was detected using an anti-US28 antibody (US28, red). Nanobody binding was detected using the mVenus-tag (Nb mV, green). Representative data of three independent experiments. b Cells were seeded in hanging droplet plates and expression of US28 (and intrabodies) were or were not induced with doxycycline. Representative data of three independent experiments. c Spheroid areas were quantified and normalized to the spheroid area of non-induced US28 negative (US28 neg) spheroid (n = 6 spheroids for non-induced and induced No Nb samples; n = 8 spheroids for non-induced Irr Nb mV samples; n = 12 spheroids for induced Irr Nb mV samples; n = 7 spheroids for non-induced VUN103 mV samples; n = 11 spheroids for induced VUN103 mV samples). d Western blot of phospho-STAT3 (p-STAT3), total STAT3, immediate early proteins (IE), intrabody-mVenus (mVenus) and actin of U251 cells (No Nb), U251cells transduced with a lentiviral vector expressing inducible non-US28 targeting intrabody-mVenus (Irr Nb mV) or VUN103-mVenus (VUN103 mV). Cells were not infected or infected with HCMV wild-type virus (WT). Representative blot shown from three independent experiments using independent biological replicates. Samples are derived from the same experiment and blots were processed in parallel. e U251 cells (No Nb), U251cells transduced with a lentiviral vector expressing inducible non-US28 targeting intrabody-mVenus (Irr Nb mV) or VUN103-mVenus (VUN103 mV)were uninfected or infected with HCMV wild-type virus (WT) or HCMV virus lacking US28 (HCMV ΔUS28). Conditioned serum of infected cells was harvested and VEGF levels were determined using ELISA (n = 3 independent experiments). All data are plotted as mean ± SD. Statistical analyses were performed using an unpaired two-tailed t-test. ns, p > 0.05. Source data are provided as a Source Data file.