Journal: Frontiers in Neurology

Article Title: Differential effects on TDP-43, piezo-2, tight-junction proteins in various brain regions following repetitive low-intensity blast overpressure

doi: 10.3389/fneur.2023.1237647

Figure Lengend Snippet: Piezo2 level following BOP. Quantification of piezo2 as measured by Western blot in (A) cortex, (B) frontal cortex, and (C) hippocampus following single and repeated blast. Values normalized to respective shams with same number of exposures. Data is expressed as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Primary antibodies included rabbit polyclonal antibody against TDP-43 (1:2000, ProteinTech cat# 10782-2-AP), rabbit polyclonal antibody against FAM38B/Piezo2 (1:2000, ProSci cat# 26–438), rabbit polyclonal antibody against GFAP (1,2000, Abcam, cat# ab7260), mouse monoclonal antibody against occludin (1,2000, ThermoFisher cat# 33–1,500), mouse monoclonal antibody against claudin-5 (1,2000, ThermoFisher cat# 35–2,500), and mouse monoclonal antibody against β-actin (1,20,000 abcam A2228).

Techniques: Western Blot

Journal: iScience

Article Title: High RIPK3 expression is associated with a higher risk of early kidney transplant failure

doi: 10.1016/j.isci.2023.107879

Figure Lengend Snippet: RIPK3 expression levels in baseline biopsies (A–E) Workflow depicting how baseline biopsies were evaluated for RIPK3 scoring and statistical analysis. From a total of 406 available biopsies, 374 were stained and evaluated within this study. 21 biopsies could not be assessed, and 11 biopsies came from transplants that succumbed to surgical complications, leading to their exclusion (B) Representative images of cortical specimens from baseline biopsies. The exact scores of the illustrated specimens with low and high RIPK3 expression are from left to right as follows: 0; 1.0; 2.34 and 3.0. Scale bars as depicted (C) Representative images of negative controls, specifically, (I) tumor-distant non-inflamed and non-fibrotic renal parenchyma from kidneys after tumor nephrectomy; (II) kidneys from end stage allograft failure with severe interstitial fibrosis and tubular atrophy; (III and IV) kidneys with membranous glomerulonephritis and nephrotic proteinuria. Scale bars as depicted (D) Scatterplot (with median reported in red) depicting the distribution of RIPK3 score across the investigated cohort (E) RIPK3 score is significantly higher in biopsies from deceased donors. Data are presented as scatterplot and in the graph the median is reported. p value from Mann-Whitney test is reported in figure.

Article Snippet: Rabbit polyclonal anti-RIPK3 , ProSci , 2283; RRID: AB_203256.

Techniques: Expressing, Staining, MANN-WHITNEY

Journal: iScience

Article Title: High RIPK3 expression is associated with a higher risk of early kidney transplant failure

doi: 10.1016/j.isci.2023.107879

Figure Lengend Snippet: Demographic and clinical characteristics of the corresponding donors and recipients of the 374 renal allografts b

Article Snippet: Rabbit polyclonal anti-RIPK3 , ProSci , 2283; RRID: AB_203256.

Techniques: Transplantation Assay

Journal: iScience

Article Title: High RIPK3 expression is associated with a higher risk of early kidney transplant failure

doi: 10.1016/j.isci.2023.107879

Figure Lengend Snippet: RIPK3 expression predicts kidney transplant failure (A) Kaplan-Meier estimates of death-censored transplant failure. Shown are estimates of the probabilities of the primary endpoint (i.e., the permanent need for dialysis after transplantation, which consists of both primary non-function (without surgical complications) and follow up end-stage transplant failure requiring the reinstitution of dialysis) comparing renal allograft baseline biopsies with a RIPK3 score of 0–2.0 (≤2) and greater than 2.0 (>2). Estimates are shown for the first year (left) and for the follow up period from year 2–5 (right). Data were censored for death-censored graft survival at the time of death with a functioning graft, at last day of detected kidney function, and either at 12 months (for one-year transplant failure) or at 60 months (for the follow up period 2–5 years). p-Values were calculated using the log rank test. (B) Kaplan-Meier estimates of non-death-censored transplant failure. Shown are estimates of the probabilities of the secondary endpoint, which was a composite of primary non-function (without surgical complications), follow-up end-stage transplant failure requiring the reinstitution of dialysis, or recipient death with a functioning allograft for renal allograft baseline biopsies, with a RIPK3 score 0 to 2.0 (≤2) and greater than 2.0 (>2). Estimates are shown for first year (left) and for the follow-up period from year 2–5 (right). Data were censored for non-death-censored graft survival at last day of detected kidney function and either at 12 months (for one-year transplant failure) or at 60 months (for the follow-up period 2–5 years. p-values were calculated using the log rank test.

Article Snippet: Rabbit polyclonal anti-RIPK3 , ProSci , 2283; RRID: AB_203256.

Techniques: Expressing, Transplantation Assay

Journal: iScience

Article Title: High RIPK3 expression is associated with a higher risk of early kidney transplant failure

doi: 10.1016/j.isci.2023.107879

Figure Lengend Snippet: Univariate Cox proportional hazards models for one-year death-censored transplant failure with hazard ratios (HR) and 95% confidence intervals (CI) for known risk factors

Article Snippet: Rabbit polyclonal anti-RIPK3 , ProSci , 2283; RRID: AB_203256.

Techniques: Transplantation Assay

Journal: iScience

Article Title: High RIPK3 expression is associated with a higher risk of early kidney transplant failure

doi: 10.1016/j.isci.2023.107879

Figure Lengend Snippet: Multivariate Cox proportional hazards models for one-year death-censored transplant failure with hazard ratios (HR) and 95% confidence intervals (CI) adjusted for donor and recipient associated risk factors

Article Snippet: Rabbit polyclonal anti-RIPK3 , ProSci , 2283; RRID: AB_203256.

Techniques: Transplantation Assay

Journal: iScience

Article Title: High RIPK3 expression is associated with a higher risk of early kidney transplant failure

doi: 10.1016/j.isci.2023.107879

Figure Lengend Snippet: Association of the RIPK3 Score with possible allograft and storage characteristics concerning organ quality

Article Snippet: Rabbit polyclonal anti-RIPK3 , ProSci , 2283; RRID: AB_203256.

Techniques:

Journal: iScience

Article Title: High RIPK3 expression is associated with a higher risk of early kidney transplant failure

doi: 10.1016/j.isci.2023.107879

Figure Lengend Snippet: RIPK3 expression and its association with acute tubular injury (A) Representative images of PAS reaction of cortical specimen with corresponding RIPK3 staining. Scale bar as depicted. (B and C) Frequency distribution of acute tubular injury (ATI) in the whole cohort. p-value from chi-square test is reported in figure (C) Frequency distribution of ATI in living and deceased donation cohorts, stratified above and below the RIPK3 score median. p value from chi-square test is reported in figure.

Article Snippet: Rabbit polyclonal anti-RIPK3 , ProSci , 2283; RRID: AB_203256.

Techniques: Expressing, Staining

Journal: iScience

Article Title: High RIPK3 expression is associated with a higher risk of early kidney transplant failure

doi: 10.1016/j.isci.2023.107879

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-RIPK3 , ProSci , 2283; RRID: AB_203256.

Techniques: Virus, Recombinant, Saline, Transfection, Purification, Plasmid Preparation, DC Protein Assay, Software, Extraction, Confocal Microscopy

Journal: Virus Research

Article Title: Construction of Fosmid-based SARS-CoV-2 replicons for antiviral drug screening and replication analyses in biosafety level 2 facilities

doi: 10.1016/j.virusres.2023.199176

Figure Lengend Snippet: Construction and characterization of the Δorf2.4 replicon and Δorf2 replicon. (A) Schema of the two new replicons. The Δorf2 replicon lacks only S (the brown box), while the Δorf2.4 replicon lacks S and E (the green box). (B) Electrophoretic image of the pCC2Fos-Δorf2.4 replicon and pCC2Fos-Δorf2 replicon digested with ZraI. In each of the migrating images, the larger bands are similar to each replicon and the smaller bands to pCC2Fos. The theoretical band sizes of the replicons are 25,927 and 26,179 bp for the Δorf2.4 replicon and Δorf2 replicon, respectively. (C) HiBiT signal kinetics of all three replicons. Huh-7 cells were electroporated with each replicon and the HiBiT signal was measured at the indicated time points. The significance of differences was assessed by a two-way ANOVA, considering p <0.05 as significant (**= p <0.01, ***= p <0.001, ****= p <0.0001). ns means Not Significant. (D) Immunofluorescent assay for viral protein detection. At 48 hpt, NSP8 was stained using the mouse anti-SARS-CoV-2 NSP8 antibody and goat anti-mouse IgG antibody conjugated with Alexa Fluor568 (red-colored cells), and the membrane was stained using the rabbit anti-SARS-CoV-2 membrane antibody and goat anti-rabbit IgG antibody conjugated with the Alexa Fluor 488 antibody (green-colored cells). The nucleus was stained by DAPI. (E) HiBiT luminescence in the supernatant of Δorf2 replicon-transfected cells. Huh-7 cells were electroporated with the Δorf2 replicon and HiBiT luminescence in its supernatant was measured at the indicated timepoints. For panels C and E, the averages and standard errors of at least 2 independent experiments with triplicate samples are shown. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Rabbit anti-SARS-CoV-2 Membrane Glycoprotein polyclonal antibodies (ProSci, Poway, California, USA) and mouse anti-SARS-CoV-2 NSP8 mAb (5A10, GeneTex, Irvine, California, USA) were used in a single mixture for the primary antibody reaction.

Techniques: Staining, Transfection