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. 2019 Sep 15;10(7):483. doi: 10.1038/s41419-019-1704-0

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© The Author(s) 2019

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 Nuclear translocation of TFEB (red) in cells of flaps in the Control, SUC and TRE groups by Immunofluorescence staining for TFEB (scale bar, 20 μm). b The percentages of TFEB translocation into nucleus in dermal layer were quantified and analyzed. c Transcripts of TFEB gene detected by real-time PCR. d and e The expressions of cytoplasmic TFEB, nuclear TFEB in the Control, SUC, TRE, TRE+Scramble control and TRE + TFEB shRNA groups as detected by Western blotting. The gels have been run under the same experimental conditions, and cropped blots are used here. f, g Optical density values of cytoplasmic TFEB and nuclear TFEB expressions in each group. h, i The expressions of VPS34, Beclin1, CTSD, SQSTM1/p62 and LC3II in the TRE, TRE+Scramble control and TRE+TFEB shRNA groups as assessed by Western blotting. And optical density values of these autophagy-related protein expressions in each group. j Transcripts of TFEB and its targeting genes, including Beclin1, VPS34, CTSD, SQSTM1/p62, and LC3 genes, were detected by real-time PCR. k, l The expressions of angiogenesis-related protein VEGF, Cadherin 5 and MMP9; oxidative stress-related protein SOD1, HO1, and eNOS and apoptosis-related protein Bax, CYC, and CASP3 in each group, were assessed by Western blotting. And optical density values of these protein expressions in each group. m Digital photographs of flaps of the TRE, TRE+Scramble control and TRE+TFEB shRNA groups on POD 3 and POD 7 (scale bar, 1 cm). n The percentages of survival area in the TRE, TRE+Scramble control and TRE+TFEB shRNA groups were quantified and analyzed. o Digital photographs of the inner side of flaps in the TRE, TRE+Scramble control and TRE+TFEB shRNA groups on POD7 (scale bar, 1 cm). p Histogram of percentage of tissue water content in each group. q Full field LDBF images of flaps in each group on POD 7 (scale bar, 1 cm). r The signal intensity of blood flow of flaps was quantified and analyzed in each group. s H&E staining to show vessels in area II of flaps in the TRE, TRE + Scramble control and TRE+TFEB shRNA groups (original magnifcation ×200; scale bar, 50 μm). t Histogram of percentage of MVDs in each group. Values are expressed as means ± SEM, n = 6 per group. *p < 0.05 and **p < 0.01, vs. Control group; ^#p < 0.05 and ^##p < 0.01, vs. SUC group; ^&p < 0.05 and ^&&p < 0.01, vs. TRE group; ^@p < 0.05 and ^@@p < 0.01, vs. TRE + Scramble control group