Journal: Acta Pharmacologica Sinica

Article Title: Curcumin attenuates brain edema in mice with intracerebral hemorrhage through inhibition of AQP4 and AQP9 expression

doi: 10.1038/aps.2015.47

Figure Lengend Snippet: Curcumin reduced the gene transcription of AQP4 and AQP9, but not AQP1. (A) AQP1, AQP4 and AQP9 mRNA levels were increased significantly at 24 h after ICH (cP<0.01 vs Sham). AQP4 and AQP9 mRNA levels were markedly decreased by curcumin (150 mg/kg) (fP<0.01 vs ICH), but no significant difference was found in AQP1 (P>0.05). Values are given as the mean±SD from 3 mice/group. (B) AQP4 and AQP9 mRNA levels were reduced by curcumin in a dose-dependent manner (fP<0.01 vs ICH). Values are given as the mean±SD from 3 mice/group.

Article Snippet: After being washed, blots were incubated overnight at 4 °C in primary antibody: AQP4 (1:500, sc-20812, Santa Cruz Biotechnology, USA), AQP9 (1:500, sc-28623, Santa Cruz Biotechnology, USA), NF-κB p65 (1:500, sc-372, Santa Cruz Biotechnology, USA), β-actin (1:1000, 8457, Cell Signaling Technology, USA), β-tubulin (1:1000, 2128, Cell Signaling Technology, USA), or Histone H3 (1:1000, 4499, Cell Signaling Technology, USA), followed by 1-h incubation with secondary antibody: Goat anti-rabbit HRP (1:2000, HA-1001-100, HangZhouHuaAn Biotechnology Co, China).

Techniques:

Journal: Acta Pharmacologica Sinica

Article Title: Curcumin attenuates brain edema in mice with intracerebral hemorrhage through inhibition of AQP4 and AQP9 expression

doi: 10.1038/aps.2015.47

Figure Lengend Snippet: Curcumin inhibited the protein expression of AQP4 and AQP9 in ICH mice. (A) AQP4 protein expression was increased significantly in the perihematomal area over 3 d after ICH, and especially in perivascular astrocyte endfeet (white arrow). Curcumin treatment (150 mg/kg) decreased AQP4 expression. The results of Western blot analysis showed that AQP4 expression was strongly induced on d 3 after ICH. Curcumin treatment inhibited AQP4 expression in a dose-dependent manner. (B) Immunoreactivity of AQP9 in astrocytes was increased significantly over 3 d after ICH (white arrow). Curcumin treatment (150 mg/kg) reduced AQP9 expression compared to that in the ICH group. AQP9 protein quantitation shown that AQP9 protein expression was markedly increased on d 3 after ICH, and down-regulated by curcumin in a dose-dependent manner. Scale bar=10 μm. Values are mean±SD from 3 mice/group. cP<0.01 vs Sham, fP<0.01 vsICH.

Article Snippet: After being washed, blots were incubated overnight at 4 °C in primary antibody: AQP4 (1:500, sc-20812, Santa Cruz Biotechnology, USA), AQP9 (1:500, sc-28623, Santa Cruz Biotechnology, USA), NF-κB p65 (1:500, sc-372, Santa Cruz Biotechnology, USA), β-actin (1:1000, 8457, Cell Signaling Technology, USA), β-tubulin (1:1000, 2128, Cell Signaling Technology, USA), or Histone H3 (1:1000, 4499, Cell Signaling Technology, USA), followed by 1-h incubation with secondary antibody: Goat anti-rabbit HRP (1:2000, HA-1001-100, HangZhouHuaAn Biotechnology Co, China).

Techniques: Expressing, Western Blot, Protein Quantitation

Journal: Acta Pharmacologica Sinica

Article Title: Curcumin attenuates brain edema in mice with intracerebral hemorrhage through inhibition of AQP4 and AQP9 expression

doi: 10.1038/aps.2015.47

Figure Lengend Snippet: Fe2+ increased the protein expression of AQP4 and AQP9 in astrocytes. Representative Western blot at 12 h after treatment with Fe2+ showed that AQP4 (A) and AQP9 (B) protein expression were up-regulated as the concentration of Fe2+ increased, reaching a peak at 50 μmol/L. Values are mean±SD from three independent experiments. cP<0.01 vs control.

Article Snippet: After being washed, blots were incubated overnight at 4 °C in primary antibody: AQP4 (1:500, sc-20812, Santa Cruz Biotechnology, USA), AQP9 (1:500, sc-28623, Santa Cruz Biotechnology, USA), NF-κB p65 (1:500, sc-372, Santa Cruz Biotechnology, USA), β-actin (1:1000, 8457, Cell Signaling Technology, USA), β-tubulin (1:1000, 2128, Cell Signaling Technology, USA), or Histone H3 (1:1000, 4499, Cell Signaling Technology, USA), followed by 1-h incubation with secondary antibody: Goat anti-rabbit HRP (1:2000, HA-1001-100, HangZhouHuaAn Biotechnology Co, China).

Techniques: Expressing, Western Blot, Concentration Assay, Control

Journal: Acta Pharmacologica Sinica

Article Title: Curcumin attenuates brain edema in mice with intracerebral hemorrhage through inhibition of AQP4 and AQP9 expression

doi: 10.1038/aps.2015.47

Figure Lengend Snippet: Curcumin or PDTC treatment inhibited AQP4 and AQP9 expression induced by Fe2+. (A) Immunofluoresence was performed at 12 h after treatment with Fe2+ (50 μmol/L), and fluorescence intensity of AQP4 was enhanced. After co-treatment with Fe2+ and curcumin (20 μmol/L) or Fe2+ and PDTC (10 μmol/L), AQP4 expression was decreased. The Western blot analysis showed that AQP4 expression was increased at 12 h after treatment with Fe2+ (50 μmol/L) and inhibited by co-treatment with Fe2+ and curcumin (20 μmol/L) or Fe2+ and PDTC (10 μmol/L). (B) The fluorescence intensity of AQP9 was increased at 12 h after treatment with Fe2+ (50 μmol/L) and inhibited by co-treatment with Fe2+ and curcumin (20 μmol/L) or Fe2+ and PDTC (10 μmol/L). AQP9 protein quantitation proved that AQP9 expression was up-regulated at 12 h after treatment with Fe2+ (50 μmol/L) and that this increase was reversed by co-treatment with Fe2+ and curcumin (20 μmol/L) or Fe2+ and PDTC (10 μmol/L). Scale bar=10 μm. Values are mean±SD from three independent experiments. cP<0.01 vs Fe2+.

Article Snippet: After being washed, blots were incubated overnight at 4 °C in primary antibody: AQP4 (1:500, sc-20812, Santa Cruz Biotechnology, USA), AQP9 (1:500, sc-28623, Santa Cruz Biotechnology, USA), NF-κB p65 (1:500, sc-372, Santa Cruz Biotechnology, USA), β-actin (1:1000, 8457, Cell Signaling Technology, USA), β-tubulin (1:1000, 2128, Cell Signaling Technology, USA), or Histone H3 (1:1000, 4499, Cell Signaling Technology, USA), followed by 1-h incubation with secondary antibody: Goat anti-rabbit HRP (1:2000, HA-1001-100, HangZhouHuaAn Biotechnology Co, China).

Techniques: Expressing, Fluorescence, Western Blot, Protein Quantitation

Journal: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society

Article Title: A Systematic Characterization of Aquaporin-9 Expression in Human Normal and Pathological Tissues.

doi: 10.1369/0022155416641028

Figure Lengend Snippet: Figure 1. Protein expression of AQP9 in normal tissues. (A) High membranous expression was observed in hepatocytes. (B) Medium level cytoplasmic expression was found in a subset of lymphoid cells in appendix (a, b). In bone marrow (C) and cells in red pulp of spleen (D), a subset of cells showed medium level cytoplasmic expression (c-f). (E) Moderate AQP9 immunoreactivity was also observed in cytoplasm of a subset of non-germinal center cells in tonsil (g, h). (F) In kidney, endothelial cells of renal glomeruli (i) displayed medium level expression, whereas renal tubules (j) showed weak expression. Scale, 100 µm.

Article Snippet: Anti-AQP9 antibodies sc-28623 and sc-74409 (Santa Cruz Biotechnology), as well as two internally generated antibodies were also tested on the immunohistochemistry platform but were evaluated as not suitable due to nonspecific staining and inconsistency with RNA-seq data (Supplementary Fig. 1).

Techniques: Expressing

Journal: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society

Article Title: A Systematic Characterization of Aquaporin-9 Expression in Human Normal and Pathological Tissues.

doi: 10.1369/0022155416641028

Figure Lengend Snippet: Figure 2. Protein expression of AQP9 in cancer tissues. High expression was observed in the membranes of tumor cells in hepatocel- lular carcinomas (A), whereas no expression was found in cholangiocarcinomas (B). Moreover, a high to medium level of membranous expression was noted in single cases of renal cancer (C) and urothelial cancer (D). Scale, 100 µm.

Article Snippet: Anti-AQP9 antibodies sc-28623 and sc-74409 (Santa Cruz Biotechnology), as well as two internally generated antibodies were also tested on the immunohistochemistry platform but were evaluated as not suitable due to nonspecific staining and inconsistency with RNA-seq data (Supplementary Fig. 1).

Techniques: Expressing

Journal: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society

Article Title: A Systematic Characterization of Aquaporin-9 Expression in Human Normal and Pathological Tissues.

doi: 10.1369/0022155416641028

Figure Lengend Snippet: Figure 3. Protein expression of AQP9 in synovial rheumatoid arthritis patients. Low cytoplasmic expression was observed in syno- vial lining (a) and mesenchymal cells (b) of two patients before Humira treatment (A and C). In one patient (B), the expression in synovial lining (c) was lost after Humira treatment, whereas the cytoplasmic expression was still observed in mesenchymal cells (d). In the other patient (D), the expression was unaltered in synovial lining (e), while no expression was found in mesenchymal cells (f). Scale, 100 µm.

Article Snippet: Anti-AQP9 antibodies sc-28623 and sc-74409 (Santa Cruz Biotechnology), as well as two internally generated antibodies were also tested on the immunohistochemistry platform but were evaluated as not suitable due to nonspecific staining and inconsistency with RNA-seq data (Supplementary Fig. 1).

Techniques: Expressing

Journal: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society

Article Title: A Systematic Characterization of Aquaporin-9 Expression in Human Normal and Pathological Tissues.

doi: 10.1369/0022155416641028

Figure Lengend Snippet: Figure 4. Protein expression of AQP9 in cell lines. (A) High expression was observed in membranes of LP-1 myeloma cells. Medium level expression in a subset of cells was found in U266/70 cells (B), whereas no or only very low expression was noted in U-87 MG cells (C) and HepG2 cells (D). Scale, 100 µm.

Article Snippet: Anti-AQP9 antibodies sc-28623 and sc-74409 (Santa Cruz Biotechnology), as well as two internally generated antibodies were also tested on the immunohistochemistry platform but were evaluated as not suitable due to nonspecific staining and inconsistency with RNA-seq data (Supplementary Fig. 1).

Techniques: Expressing