Journal: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: Enhanced large conductance K + channel activity contributes to the impaired myogenic response in the cerebral vasculature of Fawn Hooded Hypertensive rats

doi: 10.1152/ajpheart.00636.2013

Figure Lengend Snippet: Comparison of BK-α- and BK-β-subunit expression in cerebral vessel homogenates isolated from FHH and FHH.1BN rats. BK-α- and BK-β-subunits were probed on different blots. The same blots were then reprobed for β-actin. A: representative bands corresponding to the molecular weight of the BK-α-subunit (∼100 kDa), BK-β-subunit (∼25 kDa), and β-actin (∼42 kDa) in cerebral vessels obtained from FHH and FHH.1BN rats (n = 8 animals each; 3 sets of experiments). B: a comparison of the expression of α- and β-subunits. Numbers in parentheses indicate the number of samples studied.

Article Snippet: After transfer, the membrane was blocked with TBS-T buffer containing 20 mM Tris pH 7.5, 150 mM NaCl, 0.05% Tween, and a 5% blocking powder (Bio-Rad) at 4°C for 1 h. The blot was probed with primary antibody against BK α- and β-subunit [1:500 and 1:200, respectively; polyclonal rabbit Anti-K Ca 1.1, to amino acids 1184–1200 and Anti-sloβ1 (KCNMB1); Alomone Labs, Jerusalem, Israel] overnight at 4°C.

Techniques: Expressing, Isolation, Molecular Weight

Journal: Cell Death & Disease

Article Title: PAK1 regulates RUFY3-mediated gastric cancer cell migration and invasion

doi: 10.1038/cddis.2015.50

Figure Lengend Snippet: Overexpression of RUFY3 induces the formation of F-actin-enriched protrusion at the cell periphery. ( a ) GFP-RUFY3 localizes in F-actin-enriched invadopodia at the cell periphery of SGC-7901 cells. (Left panel) The living cell image acquisition was performed at 25 °C with SGC-7901 cells transfected with GFP-RUFY3 and undergoing a scratch wound assay, and GFP vector was used as a control. A representative image was shown. The white boxed areas in the left images ( × 100; scale bars, 200 μ m) are magnified in the right images ( × 600; scale bars, 24 μ m). The red boxed area in the right images shows that the cells expressing GFP-RUFY3 can localize at the periphery in a scratch area. (Right panel) Histogram showed the relative percentage of cells with actin protrusion at the migrating edge. Data are the average of at least three independent experiments with similar results, in which ~100 cells were counted (** P <0.01, compared with GFP vector). Protein expression was confirmed by western blotting assays using GFP-tagged antibody when equal glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the endogenous reference protein. ( b and c ) RUFY3 colocalizes with F-actin at the cell periphery. SGC-7901 cells were transiently transfected with pEGFP-C1 or pEGFP-RUFY3. Rhodamine-conjugated phalloidin was used to detect F-actin. After 24 h transfection, cells were fixed and permeabilized. ( b ) Images were captured using a scanning confocal fluorescence microscope and one confocal section is shown in each image. Scale bars, 10 μ m. ( c ) Histogram showed the relative percentage of colocalization cells expressing GFP-RUFY3 with F-actin at the cell periphery. The data show mean±S.E.M. (** P <0.01, compared with GFP vector), in which ~40 transfected cells were observed. ( d ) Colocalization of GFP-RIPX and myosinIIb at the cell periphery is shown by confocal microscopy. SGC-7901 cells were transiently transfected with GFP vector or GFP-RIPX. Colocalization of myosinIIb (red) with GFP-RIPX is shown by yellow fluorescence. Scale bars, 10 μ m. ( e ) Colocalization of GFP-RIPX and integrin β 5 at the cell periphery by plating cells on vitronectin is observed by confocal microscopy. SGC-7901 cells were transiently transfected with GFP vector or GFP-RIPX. Colocalization of integrin β 5 (red) with GFP-RIPX is shown by yellow fluorescence. Scale bars, 20 μ m. ( f ) Colocalization of GFP-RIPX and integrin α 3 β 1 at the cell periphery by plating cells on vitronectin is observed by confocal microscopy. Scale bars, 20 μ m

Article Snippet: The membrane was blocked with 5% nonfat dry milk in TBS-T (20 mM Tris, pH 7.4, 137 mM NaCl, 0.05% Tween-20) for 3 h at room temperature, and the proteins were probed with specific antibodies: GFP and His (GenScript Corporation, Nanjing, China), Flag (Shanghai Kangcheng), PAK1, integrin β 5 and myosinIIb (Cell Signaling), RUFY3 and vinculin (Santa Cruz), integrin α 3 β 1 (Bioss Inc. and Abcam).

Techniques: Over Expression, Transfection, Scratch Wound Assay Assay, Plasmid Preparation, Expressing, Western Blot, Fluorescence, Microscopy, Confocal Microscopy

Journal: Nutrients

Article Title: Dietary Soy Protein Isolate Attenuates Intestinal Immunoglobulin and Mucin Expression in Young Mice Compared with Casein

doi: 10.3390/nu12092739

Figure Lengend Snippet: Primers for qRT-PCR.

Article Snippet: The following antibodies were used: anti-Janus kinase-1 (JAK1) (bs-1439R, Bioss, Beijing, China), anti-phospho (p)-JAK1 (Tyr1034 + Tyr1035, bs-3238R, Bioss, China), anti- signal transducer and activator of transcription 6 (STAT6) (380957, ZEN BIO, China), anti-p-STAT6 (Tyr641, ab263947, Abcam, Cambridge, MA, USA), anti-pIgR (AF2800-SP, R&D Systems, Minneapolis, MN, USA), anti-TGFβ1 (bs-4538R, Bioss, China), and anti-β-actin (AP0060, Bioworld Technology Inc., Bloomington, MN, USA).

Techniques:

Journal: Nutrients

Article Title: Dietary Soy Protein Isolate Attenuates Intestinal Immunoglobulin and Mucin Expression in Young Mice Compared with Casein

doi: 10.3390/nu12092739

Figure Lengend Snippet: Effect of SPI on JAK1-STAT6 signaling and TGFβ levels in mouse ileum tissue. ( A ) qRT-PCR analysis of Jak1 , Stat6 , Tgfb1 , and Tgfb2 mRNA expression levels in mouse ileum tissue (normalized against Actb (β-actin); n = 8). ( B ) Western blotting analysis of JAK1, p -JAK1, STAT6, p -STAT6, pIgR, and TGFβ1 protein levels in mouse ileum tissue ( n = 6). ( C ) The statistical analyses of the Western blotting results. Results are normalized to β-actin ( n = 6). All data are presented as means ± SEM. NS, not significant ( p > 0.05); * p < 0.05; ** p < 0.01. Statistical significance was calculated using t -tests.

Article Snippet: The following antibodies were used: anti-Janus kinase-1 (JAK1) (bs-1439R, Bioss, Beijing, China), anti-phospho (p)-JAK1 (Tyr1034 + Tyr1035, bs-3238R, Bioss, China), anti- signal transducer and activator of transcription 6 (STAT6) (380957, ZEN BIO, China), anti-p-STAT6 (Tyr641, ab263947, Abcam, Cambridge, MA, USA), anti-pIgR (AF2800-SP, R&D Systems, Minneapolis, MN, USA), anti-TGFβ1 (bs-4538R, Bioss, China), and anti-β-actin (AP0060, Bioworld Technology Inc., Bloomington, MN, USA).

Techniques: Quantitative RT-PCR, Expressing, Western Blot

Journal: Oncotarget

Article Title: Structural composition of components of geoherb Moutan Cortex contributes to anti-diabetic nephropathy activity

doi: 10.18632/oncotarget.23771

Figure Lengend Snippet: Figure 1: Comparison on the down-regulation of Moutan Cortex from different regions on protein expression levels of ICAM-1,TGF-β1 and FN. HBZY-1 mesangial cells were treated with 200 μg/mL AGEs in the presence or absence of Moutan Cortex (MC) extract of 200 μg/mL. Aminoguanidine of 10 μM was used as the positive control while BSA (200 μg/mL) as blank control. (A) Western blotting was performed to compare the protein expression levels. (B–D) The grayscale scan results of ICAM-1, TGF-β1 and FN. a, Control; b, 200 μg/mL AGEs; c, Positive control aminoguanidine group; “d-m” represent MC from Anhui, Guizhou, Zhejiang, Henan, Hunan, Hebei, Sichuan, Chongqing, Shandong, Gansu. Data are expressed as means ± SD, n = 3. ###P < 0.001 vs. BSA group; *P < 0.05, **P < 0.01 and ***P < 0.001 vs. AGEs group; $P < 0.05, $$P < 0.01 and $$$P < 0.001 vs. MC from Anhui group.

Article Snippet: Rabbit anti-mouse FN, TGF-β1 and ICAM-1 monoclonal antibodies were offered by Boster Biological Engineering Co., Ltd. (Wuhan, China).

Techniques: Comparison, Expressing, Positive Control, Control, Western Blot

Journal: Oncotarget

Article Title: Structural composition of components of geoherb Moutan Cortex contributes to anti-diabetic nephropathy activity

doi: 10.18632/oncotarget.23771

Figure Lengend Snippet: Figure 2: Effect of Moutan Cortex from different regions on ICAM-1 and TGF-β1 protein expression levels in kidney of DN rats. After being treated with STZ and/or MC extract of 5g/kg or , immunohistochemistry was conducted to evaluate the expression levels of ICAM-1 (A) and TGF-β1 (B) of renal tissues. “a” represents normal control; “b” represents model group (DN rats); “c” represents Positive control 0.1 g/kg AG; “d-m” represent Gansu, Chongqing, Shangdong, Sichuan, Zhejiang, Anhui, Hunan, Guizhou, Hebei, Henan.

Article Snippet: Rabbit anti-mouse FN, TGF-β1 and ICAM-1 monoclonal antibodies were offered by Boster Biological Engineering Co., Ltd. (Wuhan, China).

Techniques: Expressing, Immunohistochemistry, Control, Positive Control