Journal: Kidney360

Article Title: Natural Killer Lymphocytes Mediate Renal Fibrosis Due to Acute Cardiorenal Syndrome

doi: 10.34067/KID.0000000000000305

Figure Lengend Snippet: Kidney mesenchymal cells expand after CA/CPR. (A) Representative flow cytometry plots of PDGFRβ+ mesenchymal cells in the kidney after CA/CPR. Plots shown are gated on live, single cells. (B) Quantification of (A). (C) Representative flow cytometry plots of CD45+PDGFRβ+ mesenchymal cells in the kidney at different time points after CA/CPR. (D) Quantification of (C). (E) Representative 20× immunofluorescence images of PDGFRα/β staining at different time points after CA/CPR. (F) Representative 20× immunofluorescence images of αSMA staining at different time points after CA/CPR. (G) Quantification of (F). (H) qRT-PCR for Acta2 (codes for αSMA) and Col1a1 (codes for type 1 collage) in the kidney at 49 days after CA/CPR compared with sham. *P < 0.05, ***P < 0.001 compared with sham in one-way ANOVA. n=4–8/group. Scale bars=100 μm. CA/CPR, cardiac arrest/cardiopulmonary resuscitation; #αSMA+, number of αSMA positive cells per high-powered field; αSMA, α smooth muscle actin; PDGFRα/β, platelet-derived growth factor α and β.

Article Snippet: Sections were then incubated with primary antibody (rabbit anti-PDGFR α / β , Abcam ab32570, 1:1000, and rat anti-CD45, BD Biosciences 550539, 1:1000) in 1% bovine serum albumin in 0.01 M PBS and 0.1% Triton X-100 at 4°C overnight.

Techniques: Flow Cytometry, Immunofluorescence, Staining, Quantitative RT-PCR, Derivative Assay

Journal: Kidney360

Article Title: Natural Killer Lymphocytes Mediate Renal Fibrosis Due to Acute Cardiorenal Syndrome

doi: 10.34067/KID.0000000000000305

Figure Lengend Snippet: Immune cells colocalize with mesenchymal cells during AKI-CKD transition. Representative 40× immunofluorescence images of different time points after CA/CPR, including 3 days (A), 7 days (B), and 49 days (C). CD45 used to label immune cells, and PDGFRα/β used to label mesenchymal cells. Scale bar=100 μm.

Article Snippet: Sections were then incubated with primary antibody (rabbit anti-PDGFR α / β , Abcam ab32570, 1:1000, and rat anti-CD45, BD Biosciences 550539, 1:1000) in 1% bovine serum albumin in 0.01 M PBS and 0.1% Triton X-100 at 4°C overnight.

Techniques: Immunofluorescence

Journal: Investigative Ophthalmology & Visual Science

Article Title: A Novel Rat Model of Dry Eye Induced by Aerosol Exposure of Particulate Matter

doi: 10.1167/iovs.63.1.39

Figure Lengend Snippet: Inflammatory cell infiltration in corneal and conjunctival tissues in the control group and PM-treated groups. ( A ) Representative images for CD45 immunofluorescent staining of the corneal, limbus, and conjunctival epithelium/stroma on day 14. Scale bar: 100 µm. There was almost no infiltration of CD45-positive cells in the central corneal tissues of the control group and the PM-treated groups, whereas a few positive cells were found in the limbus, and the numbers in the PM-treated groups increased significantly. Compared with the control group, a significantly increase in CD45-positive cells was observed in the conjunctiva of the PM-treated groups after 14 days of treatment ( B ). Each value represents the mean ± SD, n = 5. * P < 0.05 PM-high versus control, # P < 0.05 PM-high versus PM-low, v P < 0.05 PM-low versus control.

Article Snippet: Rabbit anti-rat Ki67 antibody (Servicebio, GB111141, Wuhan, China) at a 1: 800 dilution, and Rabbit anti-rat CD45 antibody (Servicebio, GB113886) at a 1: 300 dilution was used as the primary antibody, followed by incubation with ALEXA fluorophore-conjugated secondary antibodies (Invitrogen, USA) and counterstaining with Hoechst 33342 dye (0.5 g/mL; Invitrogen, USA).

Techniques: Control, Staining

Journal: Experimental and Therapeutic Medicine

Article Title: Effects of sevoflurane general anesthesia during early pregnancy on AIM2 expression in the hippocampus and parietal cortex of Sprague-Dawley offspring rats

doi: 10.3892/etm.2021.9900

Figure Lengend Snippet: Immunohistochemical analysis of the expression of (A) CD45 and (B) IL-1β in the CA1 and cortex. Scale bar, 50 µm. * P<0.05 vs. the control group; # P<0.05 vs. the S1 group. CA1, hippocampal cornu ammonis 1; cortex, parietal cortex; S1, 2-h sevoflurane general anesthesia; S2, 4-h sevoflurane general anesthesia.

Article Snippet: Rabbit anti-rat CD45 (1:1,000; cat. no. orb10328; Biorbyt Ltd.), IL-1β (1:1,000; cat. no. orb499934; Biorbyt Ltd.), caspase-1 (1:800; cat. no. PA5-86936; Thermo Fisher Scientific, Inc.), caspase-1 p10 (1:1,000; cat. no. PA5-39882; Thermo Fisher Scientific, Inc.) and β-actin (1:2,000; cat. no. orb178392; Biorbyt Ltd.) polyclonal antibodies were diluted with a TBST solution containing 3% bovine serum protein (cat. no. ab64009; Abcam) and used to probe the membranes at 4 ̊C overnight.

Techniques: Immunohistochemical staining, Expressing, Control

Journal: Experimental and Therapeutic Medicine

Article Title: Effects of sevoflurane general anesthesia during early pregnancy on AIM2 expression in the hippocampus and parietal cortex of Sprague-Dawley offspring rats

doi: 10.3892/etm.2021.9900

Figure Lengend Snippet: Western blot analysis of the protein expression levels of CD45, IL-1β pro-caspase-1 and caspase-1 p10 in the hippocampus (A) and parietal cortex (B). β-actin was used as the internal reference. * P<0.05 vs. the control group; # P<0.05 vs. the S1 group. S1, 2-h sevoflurane general anesthesia; S2, 4-h sevoflurane general anesthesia.

Article Snippet: Rabbit anti-rat CD45 (1:1,000; cat. no. orb10328; Biorbyt Ltd.), IL-1β (1:1,000; cat. no. orb499934; Biorbyt Ltd.), caspase-1 (1:800; cat. no. PA5-86936; Thermo Fisher Scientific, Inc.), caspase-1 p10 (1:1,000; cat. no. PA5-39882; Thermo Fisher Scientific, Inc.) and β-actin (1:2,000; cat. no. orb178392; Biorbyt Ltd.) polyclonal antibodies were diluted with a TBST solution containing 3% bovine serum protein (cat. no. ab64009; Abcam) and used to probe the membranes at 4 ̊C overnight.

Techniques: Western Blot, Expressing, Control