Journal: medRxiv

Article Title: Polymorphism in IFNAR contributes to glucocorticoid response and outcome in ARDS and COVID-19

doi: 10.1101/2022.03.10.22272123

Figure Lengend Snippet: (A ) STAT1 expression in the lung after 4-day culture in the presence of IFN beta with or without hydrocortisone (HC). ( B) pSTAT1 expression in the same specimens as in A. ( C) Example photomicrographs showing higher STAT2 expression in a TT patient than in a CT patient and the effect of HC on its nuclear translocation. Most STAT2 remains in the cytoplasm of the CT patients, whereas nuclear expression is prominent in the TT patient. Indicated insets are shown in the bottom row. Arrows. ( D ) Combined results of all patients noting that two CT samples are excluded in the data as the patients were already under glucocorticoid treatment at the time of sample acquisition. Ns, not significant; *P<0.05; **P<0.01; and ***P<0.001

Article Snippet: The first stage antibodies were anti-alpha chain of the IFN alpha/beta receptor (St John’s Laboratory STJ112765) that was used 1:2000 and 1:5000 and anti-Stat1 (1:400, 9175S), anti-pStat1(1:100, 9167S) and anti-Stat2 (1:200, 72604S) all from Cell Signalling.

Techniques: Expressing, Translocation Assay

Journal: Frontiers in Pharmacology

Article Title: I K1 Channel Agonist Zacopride Alleviates Cardiac Hypertrophy and Failure via Alterations in Calcium Dyshomeostasis and Electrical Remodeling in Rats

doi: 10.3389/fphar.2019.00929

Figure Lengend Snippet: Zacopride improved Iso-induced maladaptive protein alterations in rat hearts. (A) Representative western blotting images 3 days, 10 days, and 30 days after the onset of Iso infusion. (B) Dynamic changes of Kir2.1 protein expression levels. (C) Dynamic changes of p -CaMKII protein expression levels relative to total CaMKII. (D) Dynamic changes in p -RyR2 protein expression levels relative to total RyR2. (E) Dynamic changes in SERCA2 protein expression. (F) Dynamic changes of cleaved caspase 3 protein expression. All data were normalized to control. Iso, isoproterenol; Zac, zacopride; Chlo, chloroquine. Data were presented as mean ± SEM (n = 3). * P < 0.05, ** P < 0.01 compared with control. # P < 0.05, ## P < 0.01 compared with Iso+Zac.

Article Snippet: Standard western blotting were performed using respective antibodies to quantify the relative levels of Kir2.1 (mouse monoclonal anti-Kir2.1, dilution 1:1,000, Sigma; or rabbit monoclonal anti-Kir2.1, 1:1,000, Abcam), CaMKII and phosphorylated CaMKII (rabbit monoclonal anti-CaMKII, 1:1,000, Cell Signaling), SERCA2 (rabbit polyclonal anti-SERCA2, 1:1,000, Cell Signaling), RyR2-phospho S2808 or total RyR2 (rabbit polyclonal anti-( p )-RyR2, 1:1,000, Abcam), cleaved caspase 3 (rabbit monoclonal anti-c-caspase 3, 1:1,000, Cell Signaling).

Techniques: Western Blot, Expressing, Control

Journal: Brain Pathology

Article Title: Antenatal low‐intensity pulsed ultrasound reduces neurobehavioral deficits and brain injury following dexamethasone‐induced intrauterine growth restriction

doi: 10.1111/bpa.12968

Figure Lengend Snippet: Maternal LIPUS treatment enhanced the activation of CaMKII and downstream Akt signaling in IUGR brains . Representative immunoblots and quantitative data of ( A ) CaMKII, ( B ) Akt, and ( C ) ERK phosphorylation and total protein expression at PND 1 after IUGR. Representative images of double immunostaining and quantitative data of ( D ) neuron number (NeuN + ) and pAkt S473‐positive neurons, and ( E ) pCaMKII‐positive neurons at PND 1 after IUGR. Sections were stained with DAPI (blue) to visualize the nuclei. Scale bar = 50 μm. Values are mean ±standard error of mean; * p < 0.05, ** p < 0.01, and *** p < 0.001 versus saline group; # p < 0.05, ## p < 0.01, and ### p < 0.001 versus DEX group (n = 7–8/group; one‐way ANOVA). DAPI, 4'‐6‐Diamidino‐2‐phenylindole; DEX, dexamethasone; IUGR, intrauterine growth restriction; L, low‐intensity pulsed ultrasound; PND, postnatal day

Article Snippet: Protein samples obtained from brain or placenta tissue homogenates were separated on 8%–12% sodium dodecyl sulfate‐polyacrylamide gels, transferred to Immobilon‐P membranes (Millipore), and probed overnight at 4°C with primary antibodies including anti‐BDNF (1:1000; GTX132621), anti‐GDNF (1:1000; GTX17447), and anti‐VEGF (1:1000; GTX102643) from GeneTex (Irvine, CA, USA); anti‐pCaMKII (1:1000; #12716), anti‐total CaMKII (1:1000; #11945), anti‐pAkt S473 (1:1000; #9271), anti‐pAktT308 (1:1000; #4056), anti‐total Akt (1:1000; #9272), anti‐pERK (1:1000; #9101), and anti‐total ERK (1:1000; #9102) from Cell Signaling; anti‐PlGF (1:1000; ab196666), anti‐glucose transporter (GLUT) 3 (ab41525), anti‐GLUT1 (ab115730), anti‐SLC38A1 (ab134268), anti‐SNAT2 (ab90677), and anti‐SLC38A4 (ab58785) from Abcam; and anti‐β‐actin (1:10000; A5441, Sigma‐Aldrich).

Techniques: Activation Assay, Western Blot, Phospho-proteomics, Expressing, Double Immunostaining, Staining, Saline

Journal: Brain Pathology

Article Title: Antenatal low‐intensity pulsed ultrasound reduces neurobehavioral deficits and brain injury following dexamethasone‐induced intrauterine growth restriction

doi: 10.1111/bpa.12968

Figure Lengend Snippet: The schematic diagram indicates the potential mechanisms of LIPUS treatment‐induced neuroprotection in IUGR offspring brain . Our results demonstrate that antenatal LIPUS treatment promotes neuronal survival and myelination, and reduces neurodevelopmental deficits in the postnatal IUGR rats. These effects are related to increasing BDNF and PlGF expression, and subsequently enhancing neuronal CaMKII‐Akt signaling activation in the newborn IUGR brain. Meanwhile, LIPUS treatment promotes GLUT1 production and BDNF expression in the placenta. Antenatal LIPUS treatment also increases fetal weight, fetal liver weight, and placental weight after IUGR. Our findings suggest that the enhancement of cerebral BDNF‐CaMKII‐Akt signaling, and placental GLUT1 and BDNF expression by LIPUS could be a potential strategy for the management of IUGR. BDNF, brain‐derived neurotrophic factor; GLUT, glucose transporter; IUGR, intrauterine growth restriction; L, low‐intensity pulsed ultrasound; PlGF, placental growth factor.

Article Snippet: Protein samples obtained from brain or placenta tissue homogenates were separated on 8%–12% sodium dodecyl sulfate‐polyacrylamide gels, transferred to Immobilon‐P membranes (Millipore), and probed overnight at 4°C with primary antibodies including anti‐BDNF (1:1000; GTX132621), anti‐GDNF (1:1000; GTX17447), and anti‐VEGF (1:1000; GTX102643) from GeneTex (Irvine, CA, USA); anti‐pCaMKII (1:1000; #12716), anti‐total CaMKII (1:1000; #11945), anti‐pAkt S473 (1:1000; #9271), anti‐pAktT308 (1:1000; #4056), anti‐total Akt (1:1000; #9272), anti‐pERK (1:1000; #9101), and anti‐total ERK (1:1000; #9102) from Cell Signaling; anti‐PlGF (1:1000; ab196666), anti‐glucose transporter (GLUT) 3 (ab41525), anti‐GLUT1 (ab115730), anti‐SLC38A1 (ab134268), anti‐SNAT2 (ab90677), and anti‐SLC38A4 (ab58785) from Abcam; and anti‐β‐actin (1:10000; A5441, Sigma‐Aldrich).

Techniques: Expressing, Activation Assay, Derivative Assay