Journal: Cell Death & Disease

Article Title: Nerve growth factor from Chinese cobra venom stimulates chondrogenic differentiation of mesenchymal stem cells

doi: 10.1038/cddis.2017.208

Figure Lengend Snippet: Effects of NGF on BMSCs in monolayer cultures. ( a ) The MTT assay was used to detect the cytotoxic effects of NGF on BMSCs. (The values are means±S.D., n =6, * P <0.05). ( b ) The effect of NGF on the viability of BMSCs cultured in monolayers for 21 days. Scale bar, 100 μ m. ( c ) Quantification of the DNA content at 7, 14 and 21 days of culture. ( d ) HE staining was used to determine the morphology of BMCSs cultured in monolayers for 21 days. Scale bar, 100 μ m. ( e ) GAG secretion from the cells after treatment for 7, 14 and 21 days. ( f ) qRT-PCR was used to determine the expression of the ACAN , SOX9 , COL2A1 , COL1A1 , RUNX2 , ENO2, GDNF , BDNF and CNTF genes on days 7, 14 and 21. ( g and h ) Immunohistochemistry. BMSCs were stained with COL1A1 ( g ) and COL2A1 ( h ) antibodies after 21 days in culture. Scale bar, 100 μ m. The values are means±S.D., n =6; bars with different letters are significantly different from each other at P< 0.05

Article Snippet: After 1 : 200 dilutions, mouse anti-rabbit collagen type I (COL1A1, Acris OriGene Technologies, Inc., Rockville, MD, USA, TA342814) and collagen type II (COL2A1, Acris Antibodies GmbH, AF5710) antibodies were added to the cells per ections overnight.

Techniques: MTT Assay, Cell Culture, Staining, Quantitative RT-PCR, Expressing, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: Nerve growth factor from Chinese cobra venom stimulates chondrogenic differentiation of mesenchymal stem cells

doi: 10.1038/cddis.2017.208

Figure Lengend Snippet: Chondrogenic effects of NGF on BMSCs grown in 3D cultures. ( a and b ) Immunohistochemical staining of COL1A1 ( a ) and COL2A1 ( b ) was performed in the cultured constructs on days 7, 14 and 21. ( c ) qRT-PCR was performed to determine the expression levels of ACAN , SOX9 , COL2A1 , COL1A1 , RUNX2, ENO2, GDNF , BDNF and CNTF in the cultured constructs on 7 14 and 21 day. ( d ) Western blots was used to analyze the protein expression levels of Col1A1 and COL2A1. The values are means±S.D., n =6; bars with different letters are significantly different from each other at P< 0.05, Scale bar=100 μ m

Article Snippet: After 1 : 200 dilutions, mouse anti-rabbit collagen type I (COL1A1, Acris OriGene Technologies, Inc., Rockville, MD, USA, TA342814) and collagen type II (COL2A1, Acris Antibodies GmbH, AF5710) antibodies were added to the cells per ections overnight.

Techniques: Immunohistochemical staining, Staining, Cell Culture, Construct, Quantitative RT-PCR, Expressing, Western Blot

Journal: Cell Death & Disease

Article Title: Nerve growth factor from Chinese cobra venom stimulates chondrogenic differentiation of mesenchymal stem cells

doi: 10.1038/cddis.2017.208

Figure Lengend Snippet: Chondrogenic effects of NGF on BMSCs in vivo . ( a ) Safranin O staining were performed on cartilage sections. ( b and c ) Immunohistochemical staining of COL1A2 ( b ) and COL1A1 ( c ) were performed in cartilage sections. (Original low magnification × 40 (scale bar, 500 μ m); original high magnification × 200 (scale bar, 100 μ m))

Article Snippet: After 1 : 200 dilutions, mouse anti-rabbit collagen type I (COL1A1, Acris OriGene Technologies, Inc., Rockville, MD, USA, TA342814) and collagen type II (COL2A1, Acris Antibodies GmbH, AF5710) antibodies were added to the cells per ections overnight.

Techniques: In Vivo, Staining, Immunohistochemical staining

Journal: Cell Death & Disease

Article Title: Nerve growth factor from Chinese cobra venom stimulates chondrogenic differentiation of mesenchymal stem cells

doi: 10.1038/cddis.2017.208

Figure Lengend Snippet: The receptors and signaling pathways that are activated in NGF-induced BMSCs in vitro and in vivo . ( a-d ) qRT-PCR was to determine the expression levels of the ACAN , SOX9 , COL2A1 and COL1A1 genes in the repaired model after 4, 8 and 12 weeks. ( e ) Western blots was used to analyze the protein expression levels of Col1A1 and COL2A1. ( f and g ) Western blots was used to analyze the protein expression levels of the NGF receptors TrkA and p75 in vitro ( f ) and in vivo ( g ). ( h and i ) Western blots was used to analyze the protein expression of proteins in the PI3K/AKT signaling pathway, including PI3K, AKT and p-AKT, in vitro ( h ) and in vivo ( i ). ( j and k ) Western blots was used to analyze the expression of proteins in the ERK/MAPK signaling pathway, including ERK, p-ERK, P38 and p-P38, in vitro ( j ) and in vivo ( k ). ( l) Schematic description of the relevant signaling pathways that were activated by NGF. The values are means±S.D., n =10 joints; bars with different letters are significantly different from each other at P< 0.05

Article Snippet: After 1 : 200 dilutions, mouse anti-rabbit collagen type I (COL1A1, Acris OriGene Technologies, Inc., Rockville, MD, USA, TA342814) and collagen type II (COL2A1, Acris Antibodies GmbH, AF5710) antibodies were added to the cells per ections overnight.

Techniques: In Vitro, In Vivo, Quantitative RT-PCR, Expressing, Western Blot

Journal: Cell Death & Disease

Article Title: Nerve growth factor from Chinese cobra venom stimulates chondrogenic differentiation of mesenchymal stem cells

doi: 10.1038/cddis.2017.208

Figure Lengend Snippet: Primer sequences used in qRT-PCR experiments

Article Snippet: After 1 : 200 dilutions, mouse anti-rabbit collagen type I (COL1A1, Acris OriGene Technologies, Inc., Rockville, MD, USA, TA342814) and collagen type II (COL2A1, Acris Antibodies GmbH, AF5710) antibodies were added to the cells per ections overnight.

Techniques:

Journal: Toxins

Article Title: Therapeutic Targeting of Aristolochic Acid Induced Uremic Toxin Retention, SMAD 2/3 and JNK/ERK Pathways in Tubulointerstitial Fibrosis: Nephroprotective Role of Propolis in Chronic Kidney Disease

doi: 10.3390/toxins12060364

Figure Lengend Snippet: Tissue expressions of TIF, fibrotic EMT and TGF-β signaling transduction pathways among the control, PE, AAN and AAN-PE treatment groups. ( A ) C57BL/6 mice with AAN exhibited the most prominent TIF in Masson’s trichrome stain, and PE treatment ameliorated such renal injury. ( B ) PE treatment suppressed α-SMA expression and deposition of Col IaI and IV in the fibrotic EMT. ( C – I ) PE treatment attenuated not only SMAD 2/3-dependent pathways but also SMAD-independent JNK/ERK activation in the signaling cascades of TGF-β family. AAI = aristolochic acid I; AAN = aristolochic acid nephropathy; Col = collagen; TGF-β = transforming growth factor-β; PE = propolis extract; TIF = tubulointerstitial fibrosis. n = 6 in each group; ** p < 0.01 and * p < 0.05 to compare the differences between the two indicated groups.

Article Snippet: The antibodies used included anti-α-smooth muscle actin (SMA) (GeneTex, GTX100034, 1:1000, 4 °C, o/n), Col IaI (OriGene, TA309097, 1:1000, 4 °C, o/n), Col IV (Abcam, ab6586, 1:1000, 4 °C, o/n), p-Smad2/3 (Cell Signaling, 8828S, 1:1000, 4 °C, o/n), Smad2/3 (Cell Signaling, 5678S, 1:1000, 4 °C, o/n), p-JNK (Cell Signaling, 9255s, 1:1000, 4 °C, o/n), JNK (Cell Signaling, 9252, 1:1000, 4 °C, o/n), p-ERK1/2 (Cell Signaling, 9106S, 1:1000, 4 °C, o/n), ERK1/2 (Signaling, 9102, 1:1000, 4 °C, o/n) and β-actin (Santa Cruz, sc-47778, 1:2000, 4 °C, o/n).

Techniques: Transduction, Staining, Expressing, Activation Assay