Journal: Theranostics

Article Title: High OGT activity is essential for MYC-driven proliferation of prostate cancer cells

doi: 10.7150/thno.30834

Figure Lengend Snippet: Identification of transcription factors that bind to O-GlcNAc marked chromatin. A) Distribution of differently modified histone 3 reads within ±2kb of O-GlcNAc peak. B) . The top 3 motifs enriched for the O-GlcNAc ChIP-seq data showing similarity with known previously identified transcription factor motifs from published datasets. C) O-GlcNAc ChIP-seq consensus overlap with MYC ChIP-seq data reported in this study. D) O-GlcNAc ChIP-seq consensus overlap with MYC ChIP-exo dataset reported previously (GSE73994). E) OSMI-2 disrupts interaction between MYC and HCF-1. Immunoprecipitation (IP) of MYC. IgG denotes negative control. MYC was overexpressed by addition of doxycycline using the LNCaP-MYC cell line for 4 hours either in the presence or absence of 40µM OSMI-2. Experiment was repeated four times. F) Knockout of OGT disrupts the interaction between MYC and HCF-1. Experiment was performed in a mouse embryonic fibroblast cell line that has been genetically engineered to enable removal of OGT gene by addition of 0.5µM Tamoxifen (Tam). After two days of DMSO- or Tam-treatments, cell lysates were prepared and used for immunoprecipitation. Data shown is representative of two biological replicates. G) Overlap of O-GlcNAc (this study), MYC (this study) and HCF-1 (ENCSR000ECH) ChIP-seq data.

Article Snippet: Tamoxifen-inducible OGT knockout mouse embryonic fibroblast (MEF) cell line was obtained from Dr. Natasha Zachara at the CardioPEG CoreC4 (NHLBI P01 HL107153) at Johns Hopkins University School of Medicine .

Techniques: Modification, ChIP-sequencing, Immunoprecipitation, Negative Control, Knock-Out

Journal: Theranostics

Article Title: Open Porous Microenvironment-regulatory Microspheres Loaded with Curcumin@BSA NPs/BMSCs for Diabetic Wound Treatment

doi: 10.7150/thno.120285

Figure Lengend Snippet: Angiogenesis and migration assays In vitro . (A) Images representing scratch test of NIH-3T3 cells in a high-glucose environment. (B) Images representing the in vitro tube formation test and measurement of the total length of HUVECs exposed to BMSC@COPM in a high-glucose setting (scale bars: 100 μm). (C) Quantitative analysis of the scratch test results. (D) Under high-sugar conditions, BMSC@COPM promoted the proliferation of HUVECs, and cell viability was determined by a CCK-8 assay (n = 6). (E) Quantification of total length of HUVECs. (F) The representative WB images of HUVECs post-treatment with microspheres loaded with BMSCs. (G and H) Quantitative evaluation of HIF-1α and VEGF. (I) The expression of VEGF protein in HUVECs measured via ELISA. (J) In HUVECs, the levels of angiogenesis-related mRNAs (PDGF, FGF, and VEGF) were quantified using qRT-PCR. vs the control group, * P < 0.05, ** P < 0.01 and *** P < 0.001; vs the BMSC@COPM group, ## P < 0.01 and ### P < 0.001. The data shown in the figure are presented as the means ± SDs (n = 3, except D).

Article Snippet: The mouse embryonic fibroblast line NIH-3T3 and the macrophage-like cell line RAW 264.7 were donated by the medical research center of Mudanjiang Medical University.

Techniques: Migration, In Vitro, CCK-8 Assay, Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Control

Journal: Theranostics

Article Title: Open Porous Microenvironment-regulatory Microspheres Loaded with Curcumin@BSA NPs/BMSCs for Diabetic Wound Treatment

doi: 10.7150/thno.120285

Figure Lengend Snippet: Autophagy induction by BMSC@COPM enhances macrophage polarization and promotes migration and proliferation. (A) Transwell assays of NIH-3T3 cells were observed under a microscope, which were treated with BMSC@COPM in a high-glucose environment (Scale bar: 100 μm). (B) Representative WB blot of CD206 and CD86 in RAW 264.7 cells after were treated with LPS, rapamycin or chloroquine, respectively. (C) Quantitative analysis of CD206 and CD86 protein expression in (B). (D) Quantification of the transwell assay results. (E) Representative images of LC3-II and P62 protein blots in NIH-3T3 cells treated with HG and then treated with CQ (10 μM) or BMSC@COPM. (F) Statistical analysis for P62 and the LC3-II/LC3-I ratio in (E). (G) Representative images of LC3-I/II and P62 protein blots in RAW 264.7 cells, which were induced with LPS and then treated with 10 μM rapamycin (an autophagy inducer, RAPA), 10 μM CQ, or BMSC@COPM. (H) Statistical analysis protein expression related to autophagyin. vs the control group, ** P < 0.01 and *** P < 0.001; vs the LPS group and HG group, $ P < 0.05, $$ P < 0.01 and $$$ P < 0.001; vs the BMSC@COPM group, # P < 0.05, ## P < 0.01, ### P < 0.001. The data shown in the figure are presented as the means ± SDs (n = 3).

Article Snippet: The mouse embryonic fibroblast line NIH-3T3 and the macrophage-like cell line RAW 264.7 were donated by the medical research center of Mudanjiang Medical University.

Techniques: Migration, Microscopy, Expressing, Transwell Assay, Control

Journal: Theranostics

Article Title: Open Porous Microenvironment-regulatory Microspheres Loaded with Curcumin@BSA NPs/BMSCs for Diabetic Wound Treatment

doi: 10.7150/thno.120285

Figure Lengend Snippet: The relationship between autophagy and the PI3K/Akt/mTOR signaling pathway. (A) TEM detected autophagosomes in NIH-3T3 cells exposed to BMSC@COPM under high glucose conditions (Scale bar: 2 μm, the red arrows represent autophagosomes). (B) Representative immunohistochemical images of LC3-I/II and P62 in different animal (50 μM). (C) Representative images of CD206 immunofluorescence in RAW264.7 macrophages after coculture with BMSCs. (D) Quantification analysis of protein expression of LC3-I/II. (E) Statistical analysis for p62 protein expression. (F) Quantification of CD206 protein expression. (G) Representative immunohistochemical images of p-Akt, p-PI3K and p-mTOR in different groups of mice (50 μM). (H) Representative images of CD86 immunofluorescence in RAW264.7 macrophages after coculture with BMSCs. (I-K) Statistical analysis for protein levels of p-Akt, p-PI3K, p-mTOR in (G). (L) Quantification analysis of CD86 protein expression in (H). vs the control group, * P < 0.001, ** P < 0.01 and *** P < 0.001; vs the BMSC@COPM group , ## P < 0.01, ### P < 0.001. The data shown in the figure are presented as the means ± SDs (n = 3).

Article Snippet: The mouse embryonic fibroblast line NIH-3T3 and the macrophage-like cell line RAW 264.7 were donated by the medical research center of Mudanjiang Medical University.

Techniques: Immunohistochemical staining, Immunofluorescence, Expressing, Control