Journal: mSystems

Article Title: Paneth Cells Protect against Acute Pancreatitis via Modulating Gut Microbiota Dysbiosis

doi: 10.1128/msystems.01507-21

Figure Lengend Snippet: AP patients and experimental AP mice presented Paneth cell defects. (A) Histopathological changes and mean numbers of Paneth cells per crypt of duodenal mucosa specimens were assessed by H&E staining. Original magnification, ×200 ( n = 7 to 14 individuals per group). (B) Lysozyme expression (green) was assessed in Paneth cells of duodenal mucosa specimens by immunofluorescence (magnification, ×200) and lysozyme-positive/DAPI + quantification. (C to K) The mRNA expression levels of lysozyme (C), HD6 (D), HD5 (E), Reg3γ (F), Ang4 (G), sPLA2 (H), TGFβ (I), Wnt3a (J), and Lgr5 (K) were assessed. (L) Lysozyme expression (green) (magnification, ×200) and lysozyme-positive/DAPI + quantification in three AP models. (M) Mean number of Paneth cells per crypt in AP models. The data are presented as the means ± SD. ns, no significant difference; *, P ≤ 0.05.

Article Snippet: The membrane was blocked with 3% bovine serum albumin (BSA) for 1 h and incubated with primary antibodies, diluted in primary antibody dilution buffer (Epizyme Biotech, China), against Lgr5 (catalog number A10545; Abclonal, China), lysozyme (catalog number A0099; Dako, Denmark), Reg3γ (catalog number sc-377038; Santa Cruz Biotechnology, USA), Defa5 (catalog number A18208; Abclonal, China), Ang4 (catalog number sc-377497; Santa Cruz Biotechnology, USA), and sPLA2 (catalog number sc-58363; Santa Cruz Biotechnology, USA) overnight at 4°C.

Techniques: Staining, Expressing, Immunofluorescence

Journal: American Journal of Translational Research

Article Title: The oncogenic role of REG γ is exerted by activating the Wnt/β-catenin signaling pathway in osteosarcoma

doi:

Figure Lengend Snippet: siRNA and primer sequences

Article Snippet: Three small interfering RNAs (siRNAs) specifically targeting human REG γ (siRNA-REG γ) and a nonspecific negative control oligo (siRNA-NC) were purchased from GenePharma (Shanghai, China).

Techniques: Sequencing

Journal: American Journal of Translational Research

Article Title: The oncogenic role of REG γ is exerted by activating the Wnt/β-catenin signaling pathway in osteosarcoma

doi:

Figure Lengend Snippet: REG γ expression is upregulated in OS. (A-C) Expression of REG γ in OS tissues (T) and adjacent normal tissues (AT) as detected by IHC (A), WB (B) and qRT-PCR (C). (D and E) Expression of REG γ in two OS cell lines (MG-63 and SaoS-2) and a normal osteoblast cell line (hFOB1.19), as detected by WB (D) and qRT-PCR (E). Data are shown as the mean ± SD. *P<0.05.

Article Snippet: Three small interfering RNAs (siRNAs) specifically targeting human REG γ (siRNA-REG γ) and a nonspecific negative control oligo (siRNA-NC) were purchased from GenePharma (Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR

Journal: American Journal of Translational Research

Article Title: The oncogenic role of REG γ is exerted by activating the Wnt/β-catenin signaling pathway in osteosarcoma

doi:

Figure Lengend Snippet: Knockdown of REG γ in OS cell lines (MG-63 and SaoS-2) confirmed by qRT-PCR (A and B) and WB (C and D). Si-REG γ reduces the expression of REG γ at the mRNA (A and B) and protein levels (C and D) in OS cells. Compared to Si-NC, Si-REG γ-1 and Si-REG γ-2 inhibit more than 50% of REG γ expression and Si-REG γ-3 inhibits less than 50% of REG γ expression.

Article Snippet: Three small interfering RNAs (siRNAs) specifically targeting human REG γ (siRNA-REG γ) and a nonspecific negative control oligo (siRNA-NC) were purchased from GenePharma (Shanghai, China).

Techniques: Knockdown, Quantitative RT-PCR, Expressing

Journal: American Journal of Translational Research

Article Title: The oncogenic role of REG γ is exerted by activating the Wnt/β-catenin signaling pathway in osteosarcoma

doi:

Figure Lengend Snippet: REG γ depletion suppresses OS cell progression in vitro. A. Effect of Si-REG-γ-1 and Si-REG-γ-2 on OS cell growth as determined by a CCK-8 assay. B. Representative OS cell colony formation images after transfection of Si-REG γ versus Si-NC. C. Representative images of an EdU incorporation assay after transfection of Si-REG γ compared to after transfection of Si-NC. D. Apoptosis rates of MG-63 and SaoS-2 cells after transfection with Si-REG γ and Si-NC, as determined by flow cytometry. E. Representative flow cytometry analysis of the cell cycle distribution of MG-63 and SaoS-2 cells transfected with Si-REG γ and Si-NC. Data are shown as the mean ± SD. *P<0.05.

Article Snippet: Three small interfering RNAs (siRNAs) specifically targeting human REG γ (siRNA-REG γ) and a nonspecific negative control oligo (siRNA-NC) were purchased from GenePharma (Shanghai, China).

Techniques: In Vitro, CCK-8 Assay, Transfection, Flow Cytometry

Journal: American Journal of Translational Research

Article Title: The oncogenic role of REG γ is exerted by activating the Wnt/β-catenin signaling pathway in osteosarcoma

doi:

Figure Lengend Snippet: REG γ depletion inhibits OS cell migration and invasion. A and B. Representative images of the wound healing assay in OS cells after transfection. C and D. Representative images of the transwell invasion assay in MG-63 and SaoS-2 cells after transfection.

Article Snippet: Three small interfering RNAs (siRNAs) specifically targeting human REG γ (siRNA-REG γ) and a nonspecific negative control oligo (siRNA-NC) were purchased from GenePharma (Shanghai, China).

Techniques: Migration, Wound Healing Assay, Transfection, Transwell Invasion Assay

Journal: American Journal of Translational Research

Article Title: The oncogenic role of REG γ is exerted by activating the Wnt/β-catenin signaling pathway in osteosarcoma

doi:

Figure Lengend Snippet: REG γ deficiency impairs the activation of the Wnt/β-catenin signaling pathway in OS development. (A and B) Four important components of the Wnt/β-catenin signaling pathway (GSK-3β, β-catenin, c-myc and cyclin D1) as evaluated by WB in MG-63 (A) and SaoS-2 (B) cells after transfection. (C) GSK-3β level in SaoS-2 cells transfected with Si-NC or Si-REG γ-2 was detected by WB after Chx (100 µg/ml) treatment.

Article Snippet: Three small interfering RNAs (siRNAs) specifically targeting human REG γ (siRNA-REG γ) and a nonspecific negative control oligo (siRNA-NC) were purchased from GenePharma (Shanghai, China).

Techniques: Activation Assay, Transfection

Journal: American Journal of Translational Research

Article Title: The oncogenic role of REG γ is exerted by activating the Wnt/β-catenin signaling pathway in osteosarcoma

doi:

Figure Lengend Snippet: siRNA and primer sequences

Article Snippet: Transient transfection Three small interfering RNAs (siRNAs) specifically targeting human REG γ (siRNA-REG γ) and a nonspecific negative control oligo (siRNA-NC) were purchased from GenePharma (Shanghai, China).

Techniques: Sequencing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: REG γ Contributes to Regulation of Hemoglobin and Hemoglobin δ Subunit

doi: 10.1155/2017/7295319

Figure Lengend Snippet: REG γ -mediated Hb degradation in multiple hemopoietic tissues. (a) Hb levels in bone marrow, (b) liver, and (c) spleen tissues from two different REGγ +/+ and REGγ −/− mice. Protein was extracted and analyzed using Western blot against anti-Hb and anti-REG γ antibodies. β -Actin was use as internal control. The quantification analysis was conducted and shown as a graph. Left: Western blot. n = 3 represents the number of mice in each genotype. All the mice were sacrificed at age of 1-2 weeks old. Right: quantification analysis. Data are presented as means ± SEM from three independent experiments. ∗ p < 0.05 versus control.

Article Snippet: Anti-REG γ and anti- β -actin antibodies were purchased from Abmart; anti-Hb and human HBD antibodies were purchased from Santa Cruz Biotech Inc.; anti-p21 and anti-Smurf 1 antibodies were purchased from BD Biosciences Inc. (USA).

Techniques: Western Blot, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: REG γ Contributes to Regulation of Hemoglobin and Hemoglobin δ Subunit

doi: 10.1155/2017/7295319

Figure Lengend Snippet: Regulation of endogenous HBD in HeLa cells. (a) HeLa cells were transfected with HA-pSG5-HBD plasmid (1 μ g/ml) and HA-Smurf1 (1 μ g/ml) as a positive control, using Lipo2000 transfection reagent for 72 h incubation. Total protein extracts were analyzed by Western blot against anti-HA antibody. (b) CHX assay for endogenous degradation. HeLa cells transfected with HA-pSG5-HBD. After 64 h incubation, cells were treated 100 μ g/ml of CHX for 0, 2, 4, 6, and 8 h. The total protein extracts were subjected to Western blot analysis. β -Actin was used as loading control. (c) Quantification of CHX-treated Western blot results expressed as the means ± SEM. ∗ p < 0.05; ∗∗ p < 0.01 versus control.

Article Snippet: Anti-REG γ and anti- β -actin antibodies were purchased from Abmart; anti-Hb and human HBD antibodies were purchased from Santa Cruz Biotech Inc.; anti-p21 and anti-Smurf 1 antibodies were purchased from BD Biosciences Inc. (USA).

Techniques: Transfection, Plasmid Preparation, Positive Control, Incubation, Western Blot, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: REG γ Contributes to Regulation of Hemoglobin and Hemoglobin δ Subunit

doi: 10.1155/2017/7295319

Figure Lengend Snippet: REG γ -mediated degradation of HBD in HeLa cells (a). Cells were transiently transfected with HA-pSG5-HBD (2 μ g), FRT-REG γ wt (1 μ g), FRT-REG γ N151Y (1 μ g), HA-pSG5 vector (2 μ g), and FRT vector (1 μ g) using Lipo2000 transfection reagent for 72 h incubation. Protein expression was detected against anti-REG γ , anti-HBD, and anti- β -actin antibodies. Nontransfected HeLa cells were used as a control. (b) HeLa cells were transfected with pcDNA3.1-p21 (2 μ g) FRT-REG γ wt (1 μ g), FRT-REG γ N151Y (1 μ g), pcDNA3.1 vector (2 μ g), and FRT vector (1 μ g) using Lipo2000 transfection reagent for 72 h incubation. Protein expressions of indicated antibodies were determined by Western blot analysis. Nontransfected HeLa cells were used as controls. β -Actin was used as loading control. The quantification analysis was conducted and shown as a graph. Left: Western blot. Right: quantification analysis. Data is presented as means ± SEM. ∗ p < 0.05; ∗∗ p < 0.01 versus control.

Article Snippet: Anti-REG γ and anti- β -actin antibodies were purchased from Abmart; anti-Hb and human HBD antibodies were purchased from Santa Cruz Biotech Inc.; anti-p21 and anti-Smurf 1 antibodies were purchased from BD Biosciences Inc. (USA).

Techniques: Transfection, Plasmid Preparation, Incubation, Expressing, Control, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: REG γ Contributes to Regulation of Hemoglobin and Hemoglobin δ Subunit

doi: 10.1155/2017/7295319

Figure Lengend Snippet: REG γ promotes HBD degradation under oxidative stress. HeLa shN and HeLa shR cells were transfected with or without HA-pSG5-HBD plasmid. Cells were treated with a 5 mM concentration of PHZ for 0, 1, and 2 h. Oxidative stress via PHZ stimulation on HBD in the presence or absence of REG γ was measured by Western blotting against anti-HBD and anti-REG γ antibodies. β -Actin was used as internal control. (b) Quantification of HA-pSG5-HBD- and PHZ-treated Western blot results expressed as the means ± SEM. ∗∗ p < 0.01 versus control.

Article Snippet: Anti-REG γ and anti- β -actin antibodies were purchased from Abmart; anti-Hb and human HBD antibodies were purchased from Santa Cruz Biotech Inc.; anti-p21 and anti-Smurf 1 antibodies were purchased from BD Biosciences Inc. (USA).

Techniques: Transfection, Plasmid Preparation, Concentration Assay, Western Blot, Control