Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: DBMSC proliferation groups. A Group 1 consisted of DBMSC cultured alone in a complete DBMSC culture medium. B Group 2 consisted of DBMSC cultured with different concentrations (25–400 mM) of glucose in a complete DBMSC culture medium. C Group 3 consisted of DBMSC pretreated with 200 mM glucose for 72 h [200 (pre)], harvested and then re-cultured alone in a complete DBMSC culture medium. D DBMSCs were seeded in a 16-well plate (E-Plate 16). The culture plates were then placed in the xCELLigence system at 37 °C in a cell culture incubator, and DBMSC cell index was then monitored. DBMSC proliferation in response to different glucose concentrations by the xCELLigence system. As compared to untreated DBMSCs, DBMSC proliferation was unchanged at 25 mM glucose ( p > 0.05) but significantly increased at 50 and 200 mM glucose and then significantly reduced at 400 mM glucose, after 24 h in culture. E At 48 h in culture, and as compared to untreated DBMSCs, DBMSC proliferation was unchanged at 50 mM glucose ( p > 0.05) but significantly increased at 200 mM glucose and then significantly reduced at 25 and 400 mM glucose. F At 72 h in culture, and as compared to untreated DBMSCs, DBMSC proliferation significantly increased at 200 mM glucose but was significantly reduced at 25, 50 and 400 mM glucose. G - I The reversibility of DBMSC proliferation in response to glucose. DBMSCs were initially cultured with 200 mM glucose for 72 h and their proliferation was then determined using the xCELLigence system. At 24–72 h in culture, and as compared to untreated DBMSCs and DBMSC-treated with 200 mM glucose [200 (I)], the proliferation of DBMSC pretreated with 200 mM glucose [200 (pre)] significantly reduced

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Cell Culture

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: DBMSC migration groups. A Group 1 consisted of DBMSCs cultured alone in the upper chamber. B Group 2 consisted of DBMSCs cultured alone in the upper chamber while 200 mM glucose was added to the lower chamber. C Group 3 consisted of DBMSCs pre-treated with 200 mM glucose for 72 h (200(pre)) harvetsed and then re-cultured alone in the upper chamber while 200 mM glucose was added to the lower chamber. D DBMSCs were seeded in DBMSC serum free medium in the upper chamber of the CIM migration plate while DBMSC culture medium containing 30% FBS was added to the lower chambers. At 24 h, DBMSC [DB (T 200)] migration in response to 200 mM glucose significantly increased as compared to untreated DBMSCs (DB). The migration of DBMSCs pretreated with 200 mM glucose for 72 h (Pre-DB) in response to 200 mM glucose [Pre-DB (To 200)] significantly increased as compared to untreated DBMSCs (DB), but was unchanged as compared to DBMSC migrating in response to 200 mM glucose [DB (To 200)], p > 0.05. E The effect of glucose on DBMSC invasion through endothelial cells by the xCELLigence system. At 10 h, the pretreatment with 200 mM glucose for 72 h [200 (Pre)] significantly increased DBMSC invasion as compared to untreated DBMSCs and DBMSC cultured with 200 mM glucose while the addition of 200 mM glucose [200 (in)] during the invasion experiment had no significant effect on DBMSC invasion as compared to untreated DBMSCs. Each experiment was performed in triplicate and repeated with five independent DBMSC (passage 3) preparations. * p < 0.05. Bars represent standard errors

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Migration, Cell Culture

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: BMSCs were cultured with 200 mM glucose [200 (I)] and their adhesion was then determined using the xCELLigence system. As compared to untreated DBMSCs, the treatment with 200 mM glucose had no significant effect on DBMSC adhesion at 2 h in culture ( p > 0.05) while the adhesion of DBMSC pretreated with 200 mM glucose for 72 h was significantly increased at 2 h as compared to untreated DBMSCs, and DBMSC cultured with 200 mM glucose [200 (I)]. Each experiment was performed in triplicate and repeated with five independent DBMSC (passage 3) preparations. * p < 0.05. Bars represent standard errors

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Cell Culture

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: Flow cytometric analysis of DBMSC expression of immune markers. A - C The treatment with 200 mM glucose significantly increased the DBMSCs [200 (pre)] expression of ICAM-1, had no significant effect on IL-12 expression, p > 0.05, and significantly increased the expression of B7H4, as compared untreated DBMSCs. Each experiment was performed in triplicate and repeated with five independent DBMSC (passage 3) preparations. * p < 0.05. Bars represent standard errors

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Expressing

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: Glucose effects on DBMSC expression of oxidative genes with survival, anti-apoptotic, proliferation, and migration properties. DBMSCs were untreated (DBMSC) or treated with 200 mM glucose (TDBMSC) for 72 h

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Expressing, Migration

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: Glucose effects on DBMSC expression of oxidative genes with pro-oxidant and antioxidant properties. DBMSCs were untreated (DBMSC) or treated with 200 mM glucose (TDBMSC) for 72 h

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Expressing

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: Glucose increased DBMSC expression of genes with antioxidant, anti-inflammatory, anti-chemoattractant, and antimicrobial properties. DBMSCs were untreated (DBMSC) or treated with 200 mM glucose (TDBMSC) for 72 h

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Expressing, Clinical Proteomics

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: Glucose effects on DBMSC expression of oxidative genes. DBMSCs were untreated (DBMSC) or treated with 200 mM glucose (TDBMSC) for 72 h

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Expressing

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Preconditioning of Human Decidua Basalis Mesenchymal Stem/Stromal Cells with Glucose Increased Their Engraftment and Anti-diabetic Properties

doi: 10.1007/s13770-020-00239-7

Figure Lengend Snippet: Glucose effects on DBMSC expression of oxidative genes. DBMSCs were untreated (DBMSC) or treated with 200 mM glucose (TDBMSC) for 72 h

Article Snippet: DBMSCs were cultured in a complete DBMSC culture medium [DMEM-F12 medium containing 10% MSCFBS (Mesenchymal Stem Cell certified fetal bovine serum, catalogue number 12-662-011, Life Technologies, Grand Island, NY, USA), and antibiotics (100 μg/mL streptomycin and 100 U/mL penicillin)], while HUVEC were cultured in a complete endothelial cell growth medium (Catalogue number PCS-100-041TM, ATCC, Manassas, VA, USA).

Techniques: Expressing, Membrane

Journal: Nutrients

Article Title: Effects of Lingonberry ( Vaccinium vitis-idaea L.) Supplementation on Hepatic Gene Expression in High-Fat Diet Fed Mice

doi: 10.3390/nu13113693

Figure Lengend Snippet: The twenty genes with the largest negative fold change (FC) in the lingonberry supplemented high-fat diet (HF + LGB) group relative to the high-fat diet (HF) group. Mean expression levels are given as DESeq2-normalized counts. p -values are adjusted by false discovery rate (FDR).

Article Snippet: For PCR validation, RNA was transcribed to cDNA (Maxima First Strand cDNA Synthesis Kit, Thermo Fisher Scientific) and subjected to quantitative PCR with TaqMan Universal Master Mix (Thermo Fisher Scientific) and ABI Prism 7500 sequence detection system (Applied Biosystems, Foster City, CA, USA) using the following TaqMan Gene Expression assays (Thermo Fisher Scientific); Mm00432403_m1 ( Cd36 ), Mm03047343_m1 ( Cd68 ), Mm00617672_m1 ( Cidec ), Mm00444699_m1 ( Cxcl14 ), Mm00725580_s1 ( Cyp2c29 ), Mm00472168_m1 ( Cyp2c55 ), Mm00731567_m1 ( Cyp3a11 ), Mm01607174_mH ( Cyp3a59 ), Mm00487306_m1 ( Cyp46a1 ), Mm00492632_m1 ( Igfbp2 ), Mm00434228_m1 ( IL1b ), Mm00440181_m1 ( Lepr ), Mm00503358_m1 ( Mogat1 ), Mm01184322_m1 ( Pparg ), Mm04208126_mH ( Saa2 ), Mm00446229_m1 ( Slc2a2 ), Mm00443260_g1 ( Tnfa ).

Techniques: Expressing, Activity Assay, Antioxidant Activity Assay, Migration, Binding Assay, Sequencing

Journal: medRxiv

Article Title: Predictive Biomarkers for the Responsiveness of Recurrent Glioblastomas to Activated Killer Cell Immunotherapy

doi: 10.1101/2022.09.21.22280225

Figure Lengend Snippet: (A) Schematic workflow explaining the three steps needed to identify the TNFSF18, TNFSF4 , and IL12RB2 genes. (B) Heatmap of 64 genes based on an AUC > 0.85, an overall survival p -value > 0.05, and a progression-free survival p -value > 0.05. (C) A barplot of probability of response to AKC therapy in a random forest model. According to the black line of probability = 0.5, responders and non-responders were clearly separated. (D) The mRNA expression levels of TNFRSF18, TNFSF4 , and IL12RB2 in responders and non-responders are shown as boxplots.

Article Snippet: RT-qPCR quantitation was performed using TaqMan™ Gene expression assays [FAM] (Applied Biosystems, Carlsbad, CA, USA) for TNFSF4 (Hs01911853_s1), TNFRSF18 (Hs00188346_m1), and IL12RB2 (Hs00155486_m1) and TaqMan™ Gene expression Master Mix (Applied Biosystems, Foster City, CA, USA) on a Bio-Rad CFX96 Real-Time PCR Detection System (Bio-Rad).

Techniques: Expressing

Journal: medRxiv

Article Title: Predictive Biomarkers for the Responsiveness of Recurrent Glioblastomas to Activated Killer Cell Immunotherapy

doi: 10.1101/2022.09.21.22280225

Figure Lengend Snippet: ( A) Expression of TNFSF4, TNFRSF18, and IL12RB2 in T cells and U87MG cells was analyzed by flow cytometry. Left: Representative histogram plots. Gray histograms represent isotype controls. Right: Column graphs show the frequencies of TNFSF4 + , TNFRSF18 + , and IL12RB2 + cells. (B) Transwell migration assay: CFSE-stained NK cells were plated in the upper chamber of a Transwell insert with an 8-mm pore size. Cells were allowed to migrate for 8 hours toward U87MG cells with or without T cells in the lower chamber. T cells were pre-incubated with blocking antibodies to TNFSF4, TNFRSF18, and IL12RB2 for 1 hour. NK cells that migrated to the lower chamber were counted using a cell counter. The experiments were performed in triplicate.

Article Snippet: RT-qPCR quantitation was performed using TaqMan™ Gene expression assays [FAM] (Applied Biosystems, Carlsbad, CA, USA) for TNFSF4 (Hs01911853_s1), TNFRSF18 (Hs00188346_m1), and IL12RB2 (Hs00155486_m1) and TaqMan™ Gene expression Master Mix (Applied Biosystems, Foster City, CA, USA) on a Bio-Rad CFX96 Real-Time PCR Detection System (Bio-Rad).

Techniques: Expressing, Flow Cytometry, Transwell Migration Assay, Staining, Incubation, Blocking Assay

Journal: Journal of Translational Medicine

Article Title: UBC9-mediated regulation of K144 ubiquitination of Lamin A and its implications for hepatocellular carcinoma

doi: 10.1186/s12967-026-07722-0

Figure Lengend Snippet: Impact of UBC9 knockdown on HCC cells using a doxycycline-inducible system. ( A ) schematic of the doxycycline (Dox)-inducible gene repression system used to downregulate UBC9 expression. ( B ) Western blot analysis shows reduced UBC9 protein levels after 72-hour of Dox treatment. ( C ) colorimetric CCK8 proliferation assay reveals significantly decreased cell proliferation in UBC9-depleted cells over 72 hours. ( D ) scratch assay shows reduced cell migration in UBC9 knockdown cells, 48 hours post-Dox induction. ( E ) Matrigel invasion assay demonstrates a marked decrease in the invasive capacity of HCC cells upon UBC9 knockdown. ( F ) Western blot analysis and quantification confirming UBC9 knockdown efficiency in Huh-7 cells. ( G ) CCK-8 assay revealing the effect of UBC9 depletion on Huh-7 cell proliferation. ( H ) wound healing assay showing the impact of UBC9 knockdown on the migration of Huh-7 cells. ( I ) Transwell invasion assay demonstrating the effect of UBC9 knockdown on the invasion of Huh-7 cells. ( J ) in vivo xenograft tumor assay. Images of dissected tumors, tumor weights, and tumor volume growth curves from nude mice injected with control (NC) or UBC9-shRNA Hep3B cells. ns, p > 0.05; **, p < 0.01; ***, p < 0.001

Article Snippet: UBC9 shRNA and scrambled control shRNA sequences were cloned into the pTRIPZ lentiviral vector containing a tetracycline (doxycycline, Dox)-inducible system (Thermo Scientific, Waltham, MA, USA).

Techniques: Knockdown, Expressing, Western Blot, Proliferation Assay, Wound Healing Assay, Migration, Invasion Assay, CCK-8 Assay, Transwell Invasion Assay, In Vivo, Injection, Control, shRNA

Journal: Aging (Albany NY)

Article Title: Epithelial cell senescence induces pulmonary fibrosis through Nanog-mediated fibroblast activation

doi: 10.18632/aging.102613

Figure Lengend Snippet: Rapamycin could protect mice from bleomycin (BLM)-induced pulmonary fibrosis. Mice (n = 10 in each group) were intraperitoneally injected with vehicle (DMSO/PBS, 10%) or 5 mg/kg rapamycin every other day starting 7 days after administration of BLM (5 mg/kg). ( A ) Pulmonary fibrosis was determined by haematoxylin and eosin (H&E) staining. Collagen was revealed by Masson’s trichrome staining. The expression of p21 was measured by immunohistochemical analysis. ( B ) The protein levels of p16, p21, α-SMA and collagen I were detected by Western blot. The expression levels were quantified with ImageJ (n = 3). GAPDH was used as a loading control, *P < 0.05 and **P < 0.01. ( C , D ) The lung tissues were double stained with E-cadherin and p21 (C), α-SMA and collagen I (D) by immunofluorescence. The positive areas of p21 and collagen I were quantified by densitometry (n = 3), **P < 0.01.

Article Snippet: Rabbit monoclonal antibody against GAPDH was purchased from BOSTER (Wuhan, China).

Techniques: Injection, Staining, Expressing, Immunohistochemical staining, Western Blot, Control, Immunofluorescence

Journal: Aging (Albany NY)

Article Title: Epithelial cell senescence induces pulmonary fibrosis through Nanog-mediated fibroblast activation

doi: 10.18632/aging.102613

Figure Lengend Snippet: Epithelial cell senescence could induce pulmonary fibroblast activation via activating Wnt/β-catenin signalling. ( A – C ) MLE-12 cells were treated with bleomycin (BLM, 25 μg/ml) for the indicated times. (A, B) SA-β-gal staining and β-galactosidase activity measurement were performed to detect cellular senescence, * P < 0.05 and ** P < 0.01 vs. 0 h. ( C ) The protein levels of p21 and p16 were measured by Western blot. The expression levels were quantified with ImageJ (n = 3). GAPDH was used as a loading control, *P < 0.05 and **P < 0.01. ( D – I ) MLE-12 cells were pre-treated with or without BLM for 3 days. The medium was replaced by fresh medium without BLM and co-cultured with pulmonary fibroblasts for another 3 days. ( D ) The migration capacity of pulmonary fibroblasts was detected by using a wound-healing assay. Wound areas were calculated by ImageJ, **P < 0.01. ( E ) The proliferation ability of pulmonary fibroblasts was measured by EdU assay. The percentage of proliferating cells were calculated by ImageJ, **P < 0.01. ( F ) The protein levels of collagen I, vimentin and α-SMA were determined by Western blot. The expression levels were quantified with ImageJ (n = 3). GAPDH was used as a loading control, **P < 0.01. ( G ) Pulmonary fibroblasts were double stained with vimentin and collagen I by immunofluorescence. ( H ) Pulmonary fibroblasts were double stained with α-SMA and periostin by immunofluorescence. ( I ) The expression of β-catenin was measured by immunofluorescence. ( J ) MLE-12 cells were pre-treated with or without BLM for 3 days. MLE-12 cells were cultured with fresh medium without BLM for another 3 days. The supernatants were collected to culture pulmonary fibroblasts in the presence or absence of ICG-001. The expression of β-catenin, α-SMA and collagen I were examined by Western blot. The expression levels were quantified with ImageJ (n = 3). GAPDH was used as a loading control, *P < 0.05 and **P < 0.01.

Article Snippet: Rabbit monoclonal antibody against GAPDH was purchased from BOSTER (Wuhan, China).

Techniques: Activation Assay, Staining, Activity Assay, Western Blot, Expressing, Control, Cell Culture, Migration, Wound Healing Assay, EdU Assay, Immunofluorescence

Journal: Aging (Albany NY)

Article Title: Epithelial cell senescence induces pulmonary fibrosis through Nanog-mediated fibroblast activation

doi: 10.18632/aging.102613

Figure Lengend Snippet: Rapamycin could suppress epithelial cell senescence and fibroblast activation via impairing the production of SASP. ( A – E ) MLE-12 cells were treated with bleomycin (BLM), followed by treatment with or without rapamycin for 3 days. ( A ) The expression of p16 and p21 were measured by Western blot. The expression levels were quantified with ImageJ (n = 3). GAPDH was used as a loading control, **P < 0.01. ( B , C ) The protein levels of p16 and p21 were detected by immunofluorescence. ( D ) The mRNA levels of IL-1β, IL-6, IL-8 and TNF-α were determined by Q-PCR, ** P < 0.01 vs. Con and ## P < 0.01 vs. BLM. ( E , F ) MLE-12 cells were treated as in and co-cultured with pulmonary fibroblasts in fresh medium for another 3 days. ( E ) The proliferation ability of pulmonary fibroblasts were measured by EdU assay. The percentage of proliferating cells was calculated by ImageJ, **P < 0.01. ( F ) The expression of α-SMA and collagen I were detected by Western blot. The expression levels were quantified with ImageJ (n = 3). GAPDH was used as a loading control, **P < 0.01.

Article Snippet: Rabbit monoclonal antibody against GAPDH was purchased from BOSTER (Wuhan, China).

Techniques: Activation Assay, Expressing, Western Blot, Control, Immunofluorescence, Cell Culture, EdU Assay

Journal: Aging (Albany NY)

Article Title: Epithelial cell senescence induces pulmonary fibrosis through Nanog-mediated fibroblast activation

doi: 10.18632/aging.102613

Figure Lengend Snippet: Nanog silencing could suppress pulmonary fibroblast activation and impair the development of pulmonary fibrosis. ( A – C ) Pulmonary fibroblasts were transfected with LV-Nanog-siRNA and co-cultured with MLE-12 cells as in . ( A ) The mRNA levels of Nanog, Oct4 and Rex1 were measured by Q-PCR, ** P < 0.01 vs. Con and ## P < 0.01 vs. bleomycin (BLM). ( B ) The protein levels of collagen I and α-SMA were determined by Western blot. The expression levels were quantified with ImageJ (n = 3). GAPDH was used as a loading control, *P < 0.05 and **P < 0.01. ( C ) The expression of collagen I and α-SMA were further examined by immunofluorescence staining. ( D – G ) Mice were intratracheally injected with 5 × 10 8 TU/ml LV-Nanog-siRNA or negative control (NC) 7 days after administration of BLM. Mice were sacrificed on day 21 after BLM instillation. ( D ) Pulmonary fibrosis was determined by haematoxylin and eosin (H&E) staining and collagen I was revealed by Sirius Red/Fast Green staining. ( E ) The mRNA levels of Nanog, α-SMA and collagen I were determined by Q-PCR, ** P < 0.01 vs. NC and ## P < 0.01 vs. NC + BLM. ( F ) The protein levels of Nanog, collagen I and α-SMA were measured by Western blot. The expression levels were quantified with ImageJ (n = 3). GAPDH was used as a loading control, *P < 0.05 and **P < 0.01. ( G ) The expression of α-SMA and collagen I were further confirmed by immunofluorescence staining.

Article Snippet: Rabbit monoclonal antibody against GAPDH was purchased from BOSTER (Wuhan, China).

Techniques: Activation Assay, Transfection, Cell Culture, Western Blot, Expressing, Control, Immunofluorescence, Staining, Injection, Negative Control

Journal: Cell reports

Article Title: Tuberculosis exacerbates HIV-1 infection through IL-10/STAT3-dependent tunneling nanotube formation in macrophages

doi: 10.1016/j.celrep.2019.02.091

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: 3D migration assays 3D migration assays of cells in Matrigel (10–12 mg/mL, BD Biosciences) were performed as described ( Verollet et al., 2015a ) and quantified at 48h.

Techniques: Purification, Recombinant, Luciferase, Western Blot, Enzyme-linked Immunosorbent Assay, Fluorescence, Software, Imaging