Journal: Journal of Gastroenterology

Article Title: Butein inhibits ethanol-induced activation of liver stellate cells through TGF-β, NFκB, p38, and JNK signaling pathways and inhibition of oxidative stress

doi: 10.1007/s00535-012-0619-7

Figure Lengend Snippet: The effect of preincubation of HSCs with 10 μM butein on the phosphorylation of nuclear factor κB ( NFκB ), nuclear factor κB inhibitor ( ΙκΒ ), c-Jun N-terminal kinase ( JNK ), and p38 mitogen-activated protein kinase (MAPK; p38 ). The cells were preincubated with 10 μM butein for 24 h and subsequently exposed or not to 50 mM ethanol for 20 min. The relative densitometry readings (mean ± SD) from three independent experiments each with four separate cell cultures are shown in the lower panel . The upper panel shows representative blots of total ( t ) and phosphorylated ( p ) forms of NFκB, IκB, JNK, and p38 MAPK. *Significantly different from respective controls (cells not treated or treated only with butein), * p ≤ 0.05, ** p ≤ 0.001. # Statistically significant at p ≤ 0.05 in comparison to cells treated with ethanol alone (Wilcoxon test)

Article Snippet: After blocking for 15 min at room temperature with 10 % dried nonfat milk/TBS/0.1 % Tween 20, the membranes were probed overnight at +4 °C with primary antibodies (diluted in 1 % bovine serum albumin [BSA]/TBS/0.1 % Tween 20) as follows: rabbit polyclonal anti-procollagen type I (1:250; Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse monoclonal anti-α-SMA (1:1000; Sigma-Aldrich), mouse monoclonal anti-β-actin (1:4000; Sigma-Aldrich), mouse monoclonal anti-TIMP-1 (1:500; R&D Systems), rabbit monoclonal anti-TIMP-2 (1:1000; Sigma-Aldrich), rabbit polyclonal anti-MMP-13 (1:200; USBiological), rabbit monoclonal anti-total Smad3 (1:2000; Epitomics), rabbit monoclonal anti-phospho Smad3 (1:1000; Epitomics), rabbit polyclonal anti-total NFκB p65 (1:2000; Chemicon), rabbit polyclonal anti-phospho NFκB p65 (1:500; Rockland), rabbit polyclonal anti-total IκB-α (1:2000; Sigma-Aldrich), mouse monoclonal anti-phospho IκB-α (1:1000; USBiological), rabbit polyclonal anti-total JNK (1:2000; Sigma-Aldrich), rabbit polyclonal anti-phospho JNK 1/2 (1:1000; Sigma-Aldrich), rabbit polyclonal anti-total p38 (1: 10000; Sigma-Aldrich), and rabbit polyclonal anti-phospho p38 (1:1000; Sigma-Aldrich).

Techniques: Phospho-proteomics, Comparison

Journal: Journal of Gastroenterology

Article Title: Butein inhibits ethanol-induced activation of liver stellate cells through TGF-β, NFκB, p38, and JNK signaling pathways and inhibition of oxidative stress

doi: 10.1007/s00535-012-0619-7

Figure Lengend Snippet: Possible mechanisms of the antifibrotic effects of butein on hepatic stellate cells treated with ethanol. Butein attenuates oxidative stress and ? the phosphorylation of kinases p38 and JNK as well as Smad3. In effect, it downregulates ECM remodeling and the production of profibrotic proteins. ASK1 , apoptosis signal-regulating kinase 1; DLK , dual leucine zipper-bearing kinase; ECM , extracellular matrix; IκB , nuclear factor kappaB inhibitor; IKKs , IkappaB kinases; JNK , c-Jun N-terminal kinase; MAPK , mitogen-activated protein kinase; MEKK , mitogen-activated protein kinase/extracellular signal-regulated kinase kinase; MKK , mitogen-activated protein kinase kinase; MLK3 , mixed-lineage kinase 3; NFκB , nuclear factor kappaB; SAPK , stress-activated protein kinase; TAK , Triticum aestivum kinase; TRAF2 , tumor necrosis factor (TNF)-receptor-associated factor 2

Article Snippet: After blocking for 15 min at room temperature with 10 % dried nonfat milk/TBS/0.1 % Tween 20, the membranes were probed overnight at +4 °C with primary antibodies (diluted in 1 % bovine serum albumin [BSA]/TBS/0.1 % Tween 20) as follows: rabbit polyclonal anti-procollagen type I (1:250; Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse monoclonal anti-α-SMA (1:1000; Sigma-Aldrich), mouse monoclonal anti-β-actin (1:4000; Sigma-Aldrich), mouse monoclonal anti-TIMP-1 (1:500; R&D Systems), rabbit monoclonal anti-TIMP-2 (1:1000; Sigma-Aldrich), rabbit polyclonal anti-MMP-13 (1:200; USBiological), rabbit monoclonal anti-total Smad3 (1:2000; Epitomics), rabbit monoclonal anti-phospho Smad3 (1:1000; Epitomics), rabbit polyclonal anti-total NFκB p65 (1:2000; Chemicon), rabbit polyclonal anti-phospho NFκB p65 (1:500; Rockland), rabbit polyclonal anti-total IκB-α (1:2000; Sigma-Aldrich), mouse monoclonal anti-phospho IκB-α (1:1000; USBiological), rabbit polyclonal anti-total JNK (1:2000; Sigma-Aldrich), rabbit polyclonal anti-phospho JNK 1/2 (1:1000; Sigma-Aldrich), rabbit polyclonal anti-total p38 (1: 10000; Sigma-Aldrich), and rabbit polyclonal anti-phospho p38 (1:1000; Sigma-Aldrich).

Techniques: Phospho-proteomics

Journal: Cerebral Cortex (New York, NY)

Article Title: The Cortical Neuroimmune Regulator TANK Affects Emotional Processing and Enhances Alcohol Drinking: A Translational Study

doi: 10.1093/cercor/bhy341

Figure Lengend Snippet: TANK controls neuroimmune activation after alcohol and/or lipopolysaccharide (LPS) in the insular cortex of mice. (a) Images of p-NFκB-p65+immunoreactive cells (+IR) of the anterior insular cortex of tank KO and WT mice after alcohol (5 g/kg, i.g., 10 days) and LPS (3 mg/kg, i.p.) treatment (scale bar = 30 μm). (b) Quantification of p-NFκB-p65+IR cells. BioQuant image analysis shows that cell number of p-NFκB-p65+IR was significantly increased after 10 days of alcohol (i.g.) or LPS alone. Pretreatment of alcohol significantly enhances LPS-induced p-NFκB-p65 immunoreactivity in both WT and tank KO mice (ANOVA: P < 0.01). Anterior insular cortex of tank KO mice shows significantly decreased p-NFκB-p65+IR cells after alcohol and/or LPS compared with WT mice ( t -test; * P < 0.05, ** P < 0.01 vs. control treatment; §§ P < 0.01 vs. WT mice with same treatment).

Article Snippet: Following incubation in blocking solution, slides were processed in primary antibody p-NF-κB p65 (1:200; p-NFκB p65 (Ser 276) (SC-101749) rabbit polyclonal IgG, Santa Cruz Biotechnology, Inc., Dallas, TX) overnight at 4°C, and the secondary antibody, biotinylated goat anti-rabbit IgG (H + L; Vector Laboratories Inc., Burlingame, CA) was used at a 1:200 dilution for 1 h. The immune labeling was visualized using nickel-enhanced 3,3′-diaminobenzidine (DAB).

Techniques: Activation Assay, Control

Journal: The Journal of Clinical Investigation

Article Title: WNK1 promotes water homeostasis by acting as a central osmolality sensor for arginine vasopressin release

doi: 10.1172/JCI164222

Figure Lengend Snippet: ( A ) Genotyping of Wnk1 -cKO mice mediated by neuron-specific Syn1-Cre . Genomic tail-clip DNA was used for analysis. Lane 1, Wnk1 fl/+ ; Syn1-Cre ; lane 2, Wnk1 fl/fl ; lane 3, Wnk1 fl/fl ; Syn1-Cre . PCR is shown to detect WT vs. Wnk1 fl/fl locus (exon 2 and neo cassette are floxed). PCR forward primer F is located at exon 2. Reverse primers R1 and R2 are located at intron 2 and neo cassette, respectively. Note that Syn1-Cre is only active in neurons, so unexcised Wnk1 fl/fl locus is detected in tail-clip DNA. With large size neo cassette in the floxed locus, the F/R1 primer set does not amplify under the condition of PCR reaction. Additionally, PCR is shown to detect Syn1-Cre using Cre-specific primers. ( B ) Representative Western blot of WNK1 protein in WT and cKO brain regions shown relative to the kidney. Hippo, hippocampus. Cortex, cerebral cortex. ( C ) Quantitation (mean ± SEM) of 4 separate experiments, as shown in B . One WT and cKO mouse for each experiment. WNK1 was normalized to Gapdh and compared with the WT kidney (set as “1”). * P < 0.05, # P < 0.01, KO vs. WT by unpaired t test. ( D ) Atlas of brain section for immunofluorescent staining, as in E and F . OVLT (also known as vascular-organ-of-lamina-terminalis [VOLT]) is marked by a red line. 3V, third ventricle; MnPO, median preoptic nucleus; MPA, medial preoptic area. ( E and F ) Immunofluorescent staining of WNK1 in OVLT neurons colocalized with neuronal marker β-3 tubulin in WT ( E ) and cKO ( F ) mice. Scale bar: 100 μm.

Article Snippet: The following primary and secondary antibodies were used: anti-WNK1 (NB600-225 and AF2849, Novus Biologicals); anti-β3 tubulin (MAB1195, R&D Systems); p-WNK1 (pS382) antibody (SPC-1097, StressMarq); anti-Kv3.1 (NBP2-12903, Novus Biologicals), anti–GAPDH-HRP (sc-47724 HRP, Santa Cruz).

Techniques: Western Blot, Quantitation Assay, Staining, Marker

Journal: The Journal of Clinical Investigation

Article Title: WNK1 promotes water homeostasis by acting as a central osmolality sensor for arginine vasopressin release

doi: 10.1172/JCI164222

Figure Lengend Snippet: ( A ) Water intake, ( B ) urine volume, ( C ) plasma osmolality, ( D ) urine osmolality, ( E ) plasma AVP level, and ( F ) copeptin level of control (Ctrl) and cKO mice at either ad libitum water intake or after 24-hour water restriction (WR). The inset in A shows Western blotting analysis of abundance of total and phospho-WNK1 (p-WNK1) using antibody against total WNK1 and against S382 phospho-WNK1. Arrowheads indicate molecular size 250 kDa. Lysates from WT OVLT tissue at ad libitum water intake and after 24-hour water restriction were immunoprecipitated by anti-WNK1 antibody and probed by anti-WNK1 and anti-p-WNK1 antibody. Representative of 4 separate experiments. Each experiment consists of 1 mouse ad libitum and 1 mouse on water restriction. For statistical analysis was performed with 2-way repeated ANOVA with Šidák post hoc analysis; for statistical analysis of the inset in A , unpaired 2-tailed t test was performed. For bar graphs, n = 6–8 mice, as indicated in scatter plots.

Article Snippet: The following primary and secondary antibodies were used: anti-WNK1 (NB600-225 and AF2849, Novus Biologicals); anti-β3 tubulin (MAB1195, R&D Systems); p-WNK1 (pS382) antibody (SPC-1097, StressMarq); anti-Kv3.1 (NBP2-12903, Novus Biologicals), anti–GAPDH-HRP (sc-47724 HRP, Santa Cruz).

Techniques: Control, Western Blot, Immunoprecipitation

Journal: The Journal of Clinical Investigation

Article Title: WNK1 promotes water homeostasis by acting as a central osmolality sensor for arginine vasopressin release

doi: 10.1172/JCI164222

Figure Lengend Snippet: ( A ) Plasma osmolality, ( B ) [Na + ], ( C ) relative p-WNK1/WNK1 ratio in OVLT, ( D ) plasma AVP, ( E ) urine volume, ( F ) urine osmolality, and ( G ) water intake in WT and Wnk1 -cKO mice after mannitol or vehicle injection. Urine volume and water intake were measured 120 minutes after injection. Other measurements were taken 30 minutes after injection in separate mice from those in which urine and water intake were measured. The inset in C is representative of 3 experiments. Each experiment consists of 1 mouse injected with vehicle and 1 mouse injected with mannitol. Statistical analysis in A , B , and D was performed with 2-way repeated ANOVA with Šidák post hoc analysis; otherwise, unpaired t test was used. For bar graphs in A , B , D – G , n = 5 mice for each experimental condition, as indicated in scatter plot.

Article Snippet: The following primary and secondary antibodies were used: anti-WNK1 (NB600-225 and AF2849, Novus Biologicals); anti-β3 tubulin (MAB1195, R&D Systems); p-WNK1 (pS382) antibody (SPC-1097, StressMarq); anti-Kv3.1 (NBP2-12903, Novus Biologicals), anti–GAPDH-HRP (sc-47724 HRP, Santa Cruz).

Techniques: Injection

Journal: The Journal of Clinical Investigation

Article Title: WNK1 promotes water homeostasis by acting as a central osmolality sensor for arginine vasopressin release

doi: 10.1172/JCI164222

Figure Lengend Snippet: ( A ) Ruptured whole-cell current-clamp recording for membrane potentials. Pipette and bath solution are indicated. ( B and C ) Membrane potentials of freshly isolated OVLT neurons at baseline, after incubation with 5 mM NaCl for 3 minutes and 5 minutes after washout of 5 mM NaCl hypertonicity. 600 pA currents were injected to depolarize membrane potential from the resting potential –55 mV to +150 mV. B and C represent examples of NaCl-responsive and nonresponsive neurons, respectively. ( D ) Treatment with pan-WNK kinase inhibitor (WNK463). Green and cyan bars indicate responsive (R) and nonresponsive (NR), respectively. WNK463 treatment significantly decreased the percentage distribution of responsive neurons vs. vehicle (Veh) treatment. P < 0.01, WNK463 vs. Veh, by 2-tailed Fisher’s exact test. ( E ) Wnk1 -cKO eliminated NaCl responsiveness. P < 0.01, cKO vs. WT, by 2-tailed Fisher’s exact test. In D and E , OVLT neurons were isolated form 4–5 mice for vehicle-treated, WNK463-treated, WT, and cKO groups.

Article Snippet: The following primary and secondary antibodies were used: anti-WNK1 (NB600-225 and AF2849, Novus Biologicals); anti-β3 tubulin (MAB1195, R&D Systems); p-WNK1 (pS382) antibody (SPC-1097, StressMarq); anti-Kv3.1 (NBP2-12903, Novus Biologicals), anti–GAPDH-HRP (sc-47724 HRP, Santa Cruz).

Techniques: Membrane, Transferring, Isolation, Incubation, Injection

Journal: The Journal of Clinical Investigation

Article Title: WNK1 promotes water homeostasis by acting as a central osmolality sensor for arginine vasopressin release

doi: 10.1172/JCI164222

Figure Lengend Snippet: ( A ) Injection of AAV-retro-Cre virus into PVN of tdTomato-EGFP reporter mice resulted in green fluorescence in neurons of OVLT nuclei, which otherwise exhibited tomato red fluorescence. Scale bar: 200 μm. ( B ) PVN injection of AAV-retro-Cre virus into Wnk1 fl/fl mice resulted in deletion of Wnk1 in OVLT compared with control experiments with injection of AAV-retro-Cre virus into PVN of WT mice. Scale bar: 100 μm. ( C ) Urine volume, ( D ) urine osmolality, and ( E ) plasma osmolality of Wnk1 fl/fl mice before and after injection with AAV-retro-Cre virus during at libitum and after water restriction (WR). ( F ) Urine volume, ( G ) urine osmolality, and ( H ) plasma osmolality of WT mice before and after injection with AAV-retro-Cre virus. Data shown are mean ± SEM from before injection (labeled retro-AAV –) and after injection (labeled retro-AAV +). Statistical analysis by 2-way repeated ANOVA with Šidák post hoc analysis. n = 4–6 mice as indicated by scatter plots.

Article Snippet: The following primary and secondary antibodies were used: anti-WNK1 (NB600-225 and AF2849, Novus Biologicals); anti-β3 tubulin (MAB1195, R&D Systems); p-WNK1 (pS382) antibody (SPC-1097, StressMarq); anti-Kv3.1 (NBP2-12903, Novus Biologicals), anti–GAPDH-HRP (sc-47724 HRP, Santa Cruz).

Techniques: Injection, Virus, Fluorescence, Control, Labeling

Journal: The Journal of Clinical Investigation

Article Title: WNK1 promotes water homeostasis by acting as a central osmolality sensor for arginine vasopressin release

doi: 10.1172/JCI164222

Figure Lengend Snippet: ( A and B ) Cooopeptin release in Wnk1 fl/fl and control WT mice with PVN injected with AAV-retro-Cre virus. Statistical comparison was made by paired t test between ad libitum and WR. ( C and D ) In separate groups of experimental ( Wnk1 fl/fl ) and control (WT) mice, OVLT neurons were isolated for recording of membrane potential oscillation. Pie charts show distribution of neurons that exhibit membrane potential oscillation responsive and nonresponsive to HTS (5 mM NaCl). P < 0.01 between pie chart in C and D by 2-tailed Fisher’s exact test. In A and B , n = 5 Wnk1 fl/fl and WT mice per experiment, as indicated in scatter plots.

Article Snippet: The following primary and secondary antibodies were used: anti-WNK1 (NB600-225 and AF2849, Novus Biologicals); anti-β3 tubulin (MAB1195, R&D Systems); p-WNK1 (pS382) antibody (SPC-1097, StressMarq); anti-Kv3.1 (NBP2-12903, Novus Biologicals), anti–GAPDH-HRP (sc-47724 HRP, Santa Cruz).

Techniques: Control, Injection, Virus, Comparison, Isolation, Membrane

Journal: The Journal of Clinical Investigation

Article Title: WNK1 promotes water homeostasis by acting as a central osmolality sensor for arginine vasopressin release

doi: 10.1172/JCI164222

Figure Lengend Snippet: WT mice or mice heterozygous for GOF Cl – -insensitive Wnk1 -knockin ( Wnk1 -KI) allele received AAV-Cre virus injection in OVLT. ( A ) Relative abundance of phospho-OSR/SPAK (p-OSR/SPAK) in KI mice before (–) and after (+) injection, as measured by Western blotting analysis of OVLT using antibody against S373-phospho-SPAK/S325-phospho-OSR1. The inset shows representative Western blotting of 3 separate experiments. Each experiment consists of 3 replicates of WT and 3 Wnk1 -KI mice. Each data point in the bar graph represents the average of 3 replicates. Statistical analysis by unpaired t test. ( B ) Plasma AVP level, ( C ) urine volume, ( D ) urine osmolality in heterozygous Wnk1 -KI mice in which OVLT was injected with AAV-Cre virus, ( E ) plasma AVP level, ( F ) urine volume, and ( G ) urine osmolality of WT mice in which OVLT was injected with AAV-Cre virus. In B – G , n = 5 mice, as indicated in line plots. Statistical analysis by paired t test.

Article Snippet: The following primary and secondary antibodies were used: anti-WNK1 (NB600-225 and AF2849, Novus Biologicals); anti-β3 tubulin (MAB1195, R&D Systems); p-WNK1 (pS382) antibody (SPC-1097, StressMarq); anti-Kv3.1 (NBP2-12903, Novus Biologicals), anti–GAPDH-HRP (sc-47724 HRP, Santa Cruz).

Techniques: Knock-In, Virus, Injection, Western Blot

Journal: The Journal of Clinical Investigation

Article Title: WNK1 promotes water homeostasis by acting as a central osmolality sensor for arginine vasopressin release

doi: 10.1172/JCI164222

Figure Lengend Snippet: ( A ) Schematic of the virus-mediated KO of Wnk1 in PVN-projecting OVLT neurons via injection of Cre-expressing retrograde virus at the PVN region. ( B ) Representative coronal section of the mouse brain injected with Cre-expressing virus at the PVN region. Scale bar: 1 mm. ( C ) Overlay of epifluorescence and IR-DIC images showing Cre-expressing neurons in the OVLT region. Scale bar: 10 μm. A recording pipette attached to a Cre-expressing cell was illustrated. ( D ) Top: Representative traces of spontaneous firing recorded from a NaCl-R neuron (R; cyan trace) and a NaCl-NR neuron (NR; red trace) in the WT mice. Bottom: Histogram of z score from the representative NaCl-R and NaCl-NR cells. ( E ) Distribution of the Δ z score in response to 5 mM NaCl stimulation of all recorded neurons in WT mice. The dashed line indicates 0.5. ( F ) Pie chart showing distribution of NaCl-R and NaCl-NR PVN-projecting OVLT neurons in WT mice. ( G ) Top: Representative traces of spontaneous firing recorded from a NaCl-R neuron and a NaCl-NR neuron in the Wnk1 –conditional KO (cKO) mice. Bottom: Histogram of z score from the representative NaCl-R and NaCl-NR cells. ( H ) Distribution of the Δ z score in response to 5 mM NaCl stimulation of all recorded neurons in Wnk1 -cKO mice. The dashed line indicates 0.5. ( I ) Pie chart showing distribution of NaCl-R and NaCl-NR PVN-projecting OVLT neurons in Wnk1 -cKO mice. * P = 0.032, between F and I , 2-tailed Fisher’s exact test. The WT group consists of recordings of 22 cells from 13 mice; the cKO group consists of 24 cells from 11 mice.

Article Snippet: The following primary and secondary antibodies were used: anti-WNK1 (NB600-225 and AF2849, Novus Biologicals); anti-β3 tubulin (MAB1195, R&D Systems); p-WNK1 (pS382) antibody (SPC-1097, StressMarq); anti-Kv3.1 (NBP2-12903, Novus Biologicals), anti–GAPDH-HRP (sc-47724 HRP, Santa Cruz).

Techniques: Virus, Injection, Expressing, Transferring

Journal: The Journal of Clinical Investigation

Article Title: WNK1 promotes water homeostasis by acting as a central osmolality sensor for arginine vasopressin release

doi: 10.1172/JCI164222

Figure Lengend Snippet: ( A ) WNK1 exists in conformational equilibrium between chloride-bound autoinhibited dimer and chloride-free activation-competent monomer. Hyperosmolality extracts water from the cell and from the catalytic core of WNK1, which facilitates chloride unbinding, allowing autophosphorylation at S382 and be activated ( – ). WNK1 may activate Kv3.1 directly or indirectly through other intermediaries such as OSR1/SPAK. ( B ) Kv3.1 is a high-threshold voltage-gated K + channel activated by membrane depolarization to –20 mV or above ( , ). Activation of Kv3.1 shortens action potential duration, increases after hyperpolarization (AHP), and thus increases firing frequency (illustrated by red trace). Conversely, inhibition of Kv3.1 decreases firing frequency (blue trace). In support of this notion, we have found that TEA increased the action potential half-width (data not shown). ( C ) Exponential curvilinear relationship between AVP release and plasma osmolality begins at the threshold of approximately 280 mOsm/kg. WNK1 activation by cellular dehydration (Excitatory pathway; thick green line) plays an important role in AVP release by hyperosmolality. Additional mechanism(s) may be involved, at least for secretion at the basal state, which may include tonic inhibition of osmosensory neurons (Inhibitory pathway; thick solid red line). Loss of hypotonicity-mediated inhibitory pathway (thick dotted red line) may also contribute to hyperosmolality-induced AVP release. Compensation by the additional pathways may account for apparent similar AVP release defects in OVLT-selective deletion of WNK1 (by direct shRNA injection) versus neuronal deletion of WNK1. Extracellular hypertonicity may also activate WNK1 signaling cascade through molecular crowding of the protein (ref. ) (data not shown).

Article Snippet: The following primary and secondary antibodies were used: anti-WNK1 (NB600-225 and AF2849, Novus Biologicals); anti-β3 tubulin (MAB1195, R&D Systems); p-WNK1 (pS382) antibody (SPC-1097, StressMarq); anti-Kv3.1 (NBP2-12903, Novus Biologicals), anti–GAPDH-HRP (sc-47724 HRP, Santa Cruz).

Techniques: Activation Assay, Membrane, Inhibition, shRNA, Injection

Journal: Journal of Traditional Chinese Medicine

Article Title: Therapeutic effects of salidroside vs pyrrolidine dithiocarbamate against severe acute pancreatitis in rats

doi: 10.19852/j.cnki.jtcm.20210707.002

Figure Lengend Snippet: Primers for quantitative real-time polymerase chain reaction

Article Snippet: Rabbit monoclonal anti-rat Beclin-1 antibody, LC3II antibody, LAMP2 antibody, IRAK1 antibody, p-IκBα antibody, IκBα antibody, p-p65 antibody, p65 antibody and GAPDH antibody were purchased from Abcam, Cambridge, UK.

Techniques:

Journal: Journal of Traditional Chinese Medicine

Article Title: Therapeutic effects of salidroside vs pyrrolidine dithiocarbamate against severe acute pancreatitis in rats

doi: 10.19852/j.cnki.jtcm.20210707.002

Figure Lengend Snippet: A: example blots of Beclin-1, LC3Ⅱ, LAMP2, IRAK1, p-IκBα, IκBα, p-p65, p65 and GAPDH; B: relative protein level of Beclin-1; C: relative protein level of LC3Ⅱ; D: relative protein level of LAMP2; E: relative protein level of IRAK1; F: relative protein level of p-IκBα; G: relative protein level of IκBα; H: relative protein level of p-IκBα/IκBα; I: relative protein level of p-p65; J: relative protein level of p65; K: relative protein level of p-p65/p65. SAP group: the rats were treated with 0.1 mL/min speed uniformly retrograde infusion of a freshly prepared 3.5% sodium taurocholate solution (0.1 mL/100 g) into the biliopancreatic duct after laparotomy. SO group: the rats were treated with an equivalent volume of normal saline solution as the 3.5% sodium taurocholate solution in the SAP group. Sal L + S group: SAP rats were treated with low-dose (5 mg/kg) Sal; Sal M + S group: SAP rats were treated with middle-dose (10 mg/kg) Sal; Sal H + S group: SAP rats were treated with high-dose (20 mg/kg) Sal; PDTC + S group: SAP rats were treated with PDTC (100 mg/kg). LC3Ⅱ: microtubule-associated protein light chain 3II; LAMP2: lysosome associated membrane protein 2; IRAK1: interleukin-1 receptor associated kinase 1; IκBα: inhibitor α of nuclear transcription factor-κB; p-IκBα: phosphorylation of inhibitor α of nuclear transcription factor-κB; p65: nuclear transcription factor-κB 65; p-p65: phosphorylation of nuclear transcription factor-κB 65; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; SO: sham operation; Sal: salidroside; PDTC: pyrrolidine dithiocarbamate; SAP: severe acute pancreatitis. bP < 0.01 vs SO group at the same time point; aP < 0.01, fP < 0.05 vs SAP group at the same time point; cP < 0.01 24 h vs 3 h in SAP group; dP < 0.01, gP < 0.05 Sal L + S 24 h group vs Sal M + S 24 h group; eP < 0.01, hP < 0.05 Sal M+S 24 h group vs Sal H + S 24 h group.

Article Snippet: Rabbit monoclonal anti-rat Beclin-1 antibody, LC3II antibody, LAMP2 antibody, IRAK1 antibody, p-IκBα antibody, IκBα antibody, p-p65 antibody, p65 antibody and GAPDH antibody were purchased from Abcam, Cambridge, UK.

Techniques:

Journal: Journal of Traditional Chinese Medicine

Article Title: Therapeutic effects of salidroside vs pyrrolidine dithiocarbamate against severe acute pancreatitis in rats

doi: 10.19852/j.cnki.jtcm.20210707.002

Figure Lengend Snippet: Comparison of IRAK1 mRNA-GAPDH ratio, IκBα mRNA-GAPDH ratio, and p65 mRNA-GAPDH ratio in the pancreas tissue in 8 groups ($\bar{x}$s± s )

Article Snippet: Rabbit monoclonal anti-rat Beclin-1 antibody, LC3II antibody, LAMP2 antibody, IRAK1 antibody, p-IκBα antibody, IκBα antibody, p-p65 antibody, p65 antibody and GAPDH antibody were purchased from Abcam, Cambridge, UK.

Techniques:

Journal: Scientific Reports

Article Title: Overexpression of YKL40,IL-6, IL-8, TNF-α in tonsils and the role of YKL40 in childhood with obstructive sleep apnea syndrome

doi: 10.1038/s41598-024-74402-8

Figure Lengend Snippet: Involvement of p65 phosphorylation in PTLCs. a. Representative P-P65, P65 and β-actin blots in Con, rhYKL40, 3-oxo, Bay groups. b. Statistical plots between the four groups. The quantification of phosphorylated P65 was normalized by β-actin blots.( n = 6, ns P > 0.05,* P < 0.05,** P < 0.01).

Article Snippet: Phospho-NF-κB p65-S536 Rabbit pAb (polyclonal antibody)(P-P65), NF-κB p65 Rabbit mAb (monoclonal antibody) ( P65)and β-actin Rabbit mAb (β-actin )were purchased from ABclonal (Wuhan, China).

Techniques: Phospho-proteomics