Journal: The FASEB Journal

Article Title: GORAB promotes embryonic lung maturation through antagonizing AKT phosphorylation, versican expression, and mesenchymal cell migration

doi: 10.1096/fj.201902075r

Figure Lengend Snippet: FIGURE 2 Evaluation of gene expression of epithelial and mesenchymal cells in E18.5 lungs. A, B, Quantification of type I (aquaporin 5, Aqp5) and type II (prosurfactant associated protein C, proSFTPC) epithelial markers by quantitative RT-PCR (A) and western blotting (B) in control (Gorab+/+) and homozygous Gorab knockout mouse models (Gorab−/−). C, Immunofluorescence labeling of AQP5 and proSFTPC in lung of Gorab+/+ or Gorab−/− fetuses, and quantification (n > 300 cells per three pairs of littermates). Nuclei are stained with DAPI (blue). D-G, Evaluation of versican (Vcan) expression in the lung of control (Gorab+/+) and homozygous Gorab knockout mouse models (Gorab−/−) by in situ hybridization (D), immunofluorescence (E), quantitative RT-PCR (F), and western blotting (G). H, Western blotting of VCAN in primary fibroblasts isolated from E18.5 Gorab+/+ and Gorab−/− fetuses. Data are shown as means ± SD (n ≥ 3 pairs of littermates). ***P < .001, NS, not statistically significant versus corresponding controls. Scale bars, 25 µm (C, E), 50 µm (D)

Article Snippet: The following primary antibodies were used: GORAB, 1:500 (Proteintech); AQP5, 1:50 (A4979, MilliporeSigma); SFTPC, 1:250 (ab40879, Abcam); VCAN, 1:200 (ab1032, MilliporeSigma); VIM, 1:200 (ab92547, Abcam); F-actin, 1:50 (ab205, Abcam,); VCL, 1:100 (ab129002, Abcam); SCGB1A1, 1:200 (sc-365992, Santa Cruz); ACTA2, 1:500 (a2547, MilliporeSigma); CDH1, 1:100 (BD-610182, BD biosciences).

Techniques: Gene Expression, Quantitative RT-PCR, Western Blot, Control, Knock-Out, Immunofluorescence, Labeling, Staining, Expressing, In Situ Hybridization, Isolation

Journal: Investigative ophthalmology & visual science

Article Title: Aquaporin5 Deficiency Aggravates ROS/NLRP3 Inflammasome-Mediated Pyroptosis in the Lacrimal Glands.

doi: 10.1167/iovs.64.1.4

Figure Lengend Snippet: FIGURE 1. Cultivation and identification of primary LG epithelial cell. (A) The characteristic of primary LG epithelial cells of 1-month-old mice by culture. (B) Western blotting showed the expression level of AQP5, epithelial cell adhesion molecule (EpCAM) and GAPDH in primary LG epithelial cells. (C) Immunofluorescence staining showed the expression of AQP5 and EpCAM in primary LG epithelial cells (green, AQP5; EpCAM, blue, 4 ′ 6-diamidine-2-benzene index [DAPI]). (A, C) Scale bars, 200 μm.

Article Snippet: The CRISPR/Cas9 technology was used to produce AQP5−/− C57BL/6 mice by Cyagen Biosciences (Guangzhou, China).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining

Journal: Investigative ophthalmology & visual science

Article Title: Aquaporin5 Deficiency Aggravates ROS/NLRP3 Inflammasome-Mediated Pyroptosis in the Lacrimal Glands.

doi: 10.1167/iovs.64.1.4

Figure Lengend Snippet: FIGURE 2. Mitochondrial dysfunction and lipid accumulation in the AQP5−/−mouse LGs. (A) H&E staining showed morphological changes and infiltration of inflammatory cells, as indicated by black arrows. (B) ROS immunofluorescence staining of primary LG epithelial cells (green). (C) ImageJ immunofluorescence spectrum analysis showed the average ROS fluorescence intensity (n = 3 samples). Representative complete pictures of each sample were used for quantitative analysis. The number of ROS-positive cells in the AQP5+/+ mice (1 month old, 11 ± 0.577; 6 months old, 40.33 ± 0.882), in the AQP5−/−mice (1 month old, 17.67 ± 0.882; 6 months old, 49 ± 1.155). (D) JC-1 green/red ratio fluorescence staining of primary LG epithelial cells. (E) ImageJ immunofluorescence spectrum analysis showed the average fluorescence intensity of JC-1 green/red ratio (n = 3 samples). Representative complete pictures of each sample were used for quantitative analysis. (F) ORO staining of LGs (n = 3 samples). (G) ORO staining intensity analyzed by ImageJ software (n = 3 samples). Representative complete pictures of each sample were used for quantitative analysis. The neutral triglyceride intensity in the AQP5+/+ mice (1 month old, 2574 ± 11.89; 6 months old, 6681 ± 231.7), in the AQP5−/−mice (1 month old, 5348 ± 110.6; 6 months old, 20,692 ± 139.9). (H) ORO staining of primary LG epithelial cells (n = 3 samples). (I) ORO staining intensity analyzed by ImageJ software (n = 3 samples). Representative complete pictures of each sample were used for quantitative analysis. The neutral triglyceride intensity in the AQP5+/+ mice (1 month old, 2392 ± 192; 6 months old, 4482 ± 204.1), in the AQP5−/−mice (1 month old, 4467 ± 217.6; 6 months old, 9620 ± 283.6). All data are expressed as the means ± standard error of the mean. *P < 0.05, **P < 0.01, ***P < 0.001. Scale bars: (A) 40 μm, (B, D) 400 μm, (F) 40 μm, (H) 100 μm.

Article Snippet: The CRISPR/Cas9 technology was used to produce AQP5−/− C57BL/6 mice by Cyagen Biosciences (Guangzhou, China).

Techniques: Staining, Immunofluorescence, Fluorescence, Software

Journal: Investigative ophthalmology & visual science

Article Title: Aquaporin5 Deficiency Aggravates ROS/NLRP3 Inflammasome-Mediated Pyroptosis in the Lacrimal Glands.

doi: 10.1167/iovs.64.1.4

Figure Lengend Snippet: FIGURE 3. Enhanced pyroptosis in the LGs of the AQP5−/−mice owing to NLRP3 inflammasome activation. (A) TUNEL staining of LGs (green, TUNEL; blue, DAPI). (B) ImageJ and GraphPad were used to calculate the result of A (n = 3 samples). Representative complete pictures of each sample were used for quantitative analysis. (C) Immunofluorescence staining of NLRP3, caspase-1 and GSDMD in LGs (green, NLRP3, caspase-1, GSDMD; blue, DAPI). D: Western blotting showed the expression level of NLRP3, Cle-caspase-1, caspase-1, Cle-GSDMD, GSDMD, IL-1β, Cle-IL-1β, and GAPDH in LGs. (E) ImageJ and GraphPad were used to compute the result D (n = 3 samples). Data were expressed as mean ± standard error of the mean. *P < 0.05, **P < 0.01, ***P < 0.001. Two-way ANOVA. Scale bars: (A) 100 μm, (C) 200 μm.

Article Snippet: The CRISPR/Cas9 technology was used to produce AQP5−/− C57BL/6 mice by Cyagen Biosciences (Guangzhou, China).

Techniques: Activation Assay, TUNEL Assay, Staining, Immunofluorescence, Western Blot, Expressing

Journal: Investigative ophthalmology & visual science

Article Title: Aquaporin5 Deficiency Aggravates ROS/NLRP3 Inflammasome-Mediated Pyroptosis in the Lacrimal Glands.

doi: 10.1167/iovs.64.1.4

Figure Lengend Snippet: FIGURE 4. Enhanced pyroptosis in the primary LG epithelial cells of the AQP5−/−mice owing to NLRP3 inflammasome activation. (A) TUNEL staining of primary LG epithelial cells (green, TUNEL; blue, DAPI). (B) ImageJ and GraphPad were used to calculate the result of A (n = 3 samples). Representative complete pictures of each sample were used for quantitative analysis. (C) Immunofluorescence staining of NLRP3, caspase-1 and GSDMD (green, NLRP3, caspase-1, GSDMD; blue, DAPI). (D) Western blotting showed the expression level of NLRP3, Cle-caspase-1, caspase-1, Cle-GSDMD, GSDMD, IL-1β, Cle-IL-1β, and GAPDH in primary LG epithelial cells. (E) ImageJ and Graph- Pad were used to compute the result D (n = 3 samples). Data were expressed as mean ± standard error of the mean. *P < 0.05, **P < 0.01, ***P < 0.001. Two-way ANOVA. Scale bars: (A) 400 μm, (C) 100 μm.

Article Snippet: The CRISPR/Cas9 technology was used to produce AQP5−/− C57BL/6 mice by Cyagen Biosciences (Guangzhou, China).

Techniques: Activation Assay, TUNEL Assay, Staining, Immunofluorescence, Western Blot, Expressing

Journal: Investigative ophthalmology & visual science

Article Title: Aquaporin5 Deficiency Aggravates ROS/NLRP3 Inflammasome-Mediated Pyroptosis in the Lacrimal Glands.

doi: 10.1167/iovs.64.1.4

Figure Lengend Snippet: FIGURE 5. LG pyroptosis reduction via NLRP3 inflammasome inhibition. (A) Sodium fluorescein staining in the cornea. (B) The lacrimal secretion of mice was measured using the phenol red cotton thread method. (C) H&E staining showed morphological changes and infiltration of inflammatory cells. (D) ORO staining of LGs (n = 3 samples). (E) ORO staining intensity analyzed by ImageJ software (n = 3 samples). Representative complete pictures of each sample were used for quantitative analysis. The neutral triglyceride intensity in the AQP5+/+ mice was 29,303 ± 2000, in the MCC950-treated AQP5+/+ mice 11,327 ± 757.9, in the AQP5−/−mice 38,206 ± 2454, and in the MCC950-treated AQP5−/−mice 16,888 ± 845.7. (F) TUNEL staining of LGs (green, TUNEL; blue, DAPI). (G) ImageJ and GraphPad were used to calculate the result F (n = 3 samples). Representative complete pictures of each sample were used for quantitative analysis. (H) Western blotting showed the expression level of NLRP3, Cle-caspase-1, caspase-1, Cle-GSDMD, GSDMD, IL-1β, Cle-IL-1β, and GAPDH in LGs. (I) ImageJ and GraphPad were used to compute the result of H (n = 3 samples). Data were expressed as mean ± standard error of the mean. *P < 0.05, **P < 0.01, ***P < 0.001. Two-way ANOVA. Scale bars: (C, D) 40 μm, (F) 100 μm.

Article Snippet: The CRISPR/Cas9 technology was used to produce AQP5−/− C57BL/6 mice by Cyagen Biosciences (Guangzhou, China).

Techniques: Inhibition, Staining, Software, TUNEL Assay, Western Blot, Expressing

Journal: Investigative ophthalmology & visual science

Article Title: Aquaporin5 Deficiency Aggravates ROS/NLRP3 Inflammasome-Mediated Pyroptosis in the Lacrimal Glands.

doi: 10.1167/iovs.64.1.4

Figure Lengend Snippet: FIGURE 6. LG pyroptosis reduction via NLRP3 inflammasome inhibition. (A) ORO staining of primary LG epithelial cells (n = 3 samples). (B) ORO staining intensity analyzed by ImageJ software (n = 3 samples). Representative complete pictures of each sample were used for quantitative analysis. The neutral triglyceride intensity in the AQP5+/+ mice was 3370.0 ± 248.2, in the MCC950-treated AQP5+/+ mice 1269 ± 101.9, in the AQP5−/−mice 23,150 ± 2350, and in the MCC950-treated AQP5−/−mice 3695 ± 164 (n = 3 samples ). (C) JC-1 green/red ratio fluorescence staining of primary LG epithelial cells. (D) ImageJ immunofluorescence spectrum analysis showed the average fluorescence intensity of JC-1 green/red ratio (n = 3 samples). Representative complete pictures of each sample were used for quantitative analysis. (E) TUNEL staining of primary LG epithelial cells (green, TUNEL; blue, DAPI). (F) ImageJ and GraphPad were used to calculate the result E (n = 3 samples). Representative complete pictures of each sample were used for quantitative analysis. (G) Western blotting showed the expression level of NLRP3, Cle-caspase-1, caspase-1, Cle-GSDMD, GSDMD, IL-1β, Cle-IL-1β and GAPDH in primary LG epithelial cells. (H) ImageJ and GraphPad were used to compute the result G (n = 3 samples). Data were expressed as mean ± standard error of the mean. *P < 0.05, **P < 0.01, ***P < 0.001. Two-way ANOVA. Scale bars: (A) 100 μm, (C, E) 400 μm.

Article Snippet: The CRISPR/Cas9 technology was used to produce AQP5−/− C57BL/6 mice by Cyagen Biosciences (Guangzhou, China).

Techniques: Inhibition, Staining, Software, Fluorescence, Immunofluorescence, TUNEL Assay, Western Blot, Expressing

Journal: Investigative ophthalmology & visual science

Article Title: Aquaporin5 Deficiency Aggravates ROS/NLRP3 Inflammasome-Mediated Pyroptosis in the Lacrimal Glands.

doi: 10.1167/iovs.64.1.4

Figure Lengend Snippet: FIGURE 7. ROS removal decreased LG pyroptosis by inhibiting NLRP3 inflammasome activation. (A) JC-1 green/red ratio fluorescence staining of primary LG epithelial cells. (B) ImageJ immunofluorescence spectrum analysis showed the average fluorescence intensity of JC-1 green/red ratio (n = 3 samples). Representative complete pictures of each sample were used for quantitative analysis. (C) TUNEL staining of primary LG epithelial cells (green, TUNEL; blue, DAPI). (D) ImageJ and GraphPad were used to calculate the result of C (n = 3 samples). Representative complete pictures of each sample were used for quantitative analysis. (E) ROS immunofluorescence staining of primary LG epithelial cells (green). (F) ImageJ immunofluorescence spectrum analysis showed the average ROS fluorescence intensity (n = 3 samples). Representative complete pictures of each sample were used for quantitative analysis. The number of ROS-positive cells in the AQP5−/−mice 52 ± 0.577 in the NAC-treated AQP5−/−mice 5.333 ± 1.202. (G) Western blotting showed the expression level of NLRP3, Cle-caspase-1, caspase-1, Cle-GSDMD, GSDMD, IL-1β, Cle-IL-1β, and GAPDH in primary LG epithelial cells. (H) ImageJ and GraphPad were used to compute the result of G (n = 3 samples). Data were expressed as mean ± standard error of the mean. *P < 0.05, **P < 0.01, ***P < 0.001. Two-way ANOVA. Scale bars: (A) 200 μm, (C, E) 100 μm.

Article Snippet: The CRISPR/Cas9 technology was used to produce AQP5−/− C57BL/6 mice by Cyagen Biosciences (Guangzhou, China).

Techniques: Activation Assay, Fluorescence, Staining, Immunofluorescence, TUNEL Assay, Western Blot, Expressing

Journal: Investigative ophthalmology & visual science

Article Title: Aquaporin5 Deficiency Aggravates ROS/NLRP3 Inflammasome-Mediated Pyroptosis in the Lacrimal Glands.

doi: 10.1167/iovs.64.1.4

Figure Lengend Snippet: FIGURE 8. Modes of the AQP5/ROS/NLRP3 inflammasome axis leading to LGs pyroptosis.

Article Snippet: The CRISPR/Cas9 technology was used to produce AQP5−/− C57BL/6 mice by Cyagen Biosciences (Guangzhou, China).

Techniques:

Journal: The Saudi Dental Journal

Article Title: Role of Mesenchymal Stem Cell Exosomes and Injectable Platelet Rich Fibrin on Structure and Function of Submandibular Salivary Gland of Aged Albino Rats

doi: 10.1007/s44445-025-00056-5

Figure Lengend Snippet: Photomicrographs of SMG showing positive cytoplasmic immunoreaction in striated duct and GCT cells in ( a ) Group 1, ( b ) Group 2, ( c ) Group 3, ( d ) group 4 (Anti-AQP5 antibody, original magnification x400, scale bar 50 µm). Bar charts representing mean and SD values of: ( e )- AQP5 area% and ( f )- NGF level of the different groups. (* Significant; ** Highly significant; NS: non-significant)

Article Snippet: Sections were then incubated with AQP5 antibodies (catalog No. E-AB-15511, Elabscience®, USA) at concentration 1:50 overnight at 4°C.

Techniques: