Journal: Cell biochemistry and function

Article Title: POFUT1 and PLAGL2 are characteristic markers of mucinous colorectal cancer associated with MUC2 expression.

doi: 10.1002/cbf.3989

Figure Lengend Snippet: FIGURE 4 Effect of POFUT1 and PLAGL2 knockdown on MUC2 expression in WiDr cells. (A) Comparison of POFUT1 and PLAGL2 expression by qRT‐PCR between control and knockdown WiDr cells (n = 3). (B) Comparison of MUC2 and KRT20 expression by qRT‐PCR between control and knockdown WiDr cells (n = 3). (C) Western blot analysis comparing the protein expression of POFUT1, PLAGL2, and MUC2 between control and knocked‐down WiDr cells. β‐actin was analyzed as a loading control. **p < .01; *p < .05.

Article Snippet: The primary antibody against POFUT1 (14929‐1‐AP; Proteintech), PLAGL2 (11540‐1‐ AP; Proteintech), MUC2 (27675‐1‐AP; Proteintech), and β‐actin (ab8227; Abcam) were diluted 1:4000, 1:2500, and 1:1000 respectively, and applied overnight at 4°C.

Techniques: Knockdown, Expressing, Comparison, Quantitative RT-PCR, Control, Western Blot

Journal: Cell biochemistry and function

Article Title: POFUT1 and PLAGL2 are characteristic markers of mucinous colorectal cancer associated with MUC2 expression.

doi: 10.1002/cbf.3989

Figure Lengend Snippet: FIGURE 5 Effect of POFUT1 and PLAGL2 knockdown in vivo. (A) Images of the tumors generated from control and knocked down WiDr cells. Scale bar, 5 mm. (B) Comparison of tumor volumes between control and knocked‐down WiDr‐derived tumors (n = 3). (C) Comparison of MUC2 expression by qRT‐PCR between control and knocked down WiDr‐derived tumors (n = 3). (D) Representative images of IHC on MUC2 in control and knocked down WiDr‐derived tumors. Scale bar, 250 μm. *p < .05.

Article Snippet: The primary antibody against POFUT1 (14929‐1‐AP; Proteintech), PLAGL2 (11540‐1‐ AP; Proteintech), MUC2 (27675‐1‐AP; Proteintech), and β‐actin (ab8227; Abcam) were diluted 1:4000, 1:2500, and 1:1000 respectively, and applied overnight at 4°C.

Techniques: Knockdown, In Vivo, Generated, Control, Comparison, Derivative Assay, Expressing, Quantitative RT-PCR

Journal: Cell biochemistry and function

Article Title: POFUT1 and PLAGL2 are characteristic markers of mucinous colorectal cancer associated with MUC2 expression.

doi: 10.1002/cbf.3989

Figure Lengend Snippet: FIGURE 6 Expression of POFUT1 and PLAGL2 in human CRCs. (A) Representative images of IHC on POFUT1, PLAGL2, and MUC2 in human MAC and NMAC tissues. Scale bar, 200 μm (B) Comparison of IHC scores between MAC and NMAC tissues. (n = 15 each). **p < .01; *p < .05. CRC, colorectal cancer; IHC, immunohistochemistry; MAC, mucinous adenocarcinoma; NMAC, nonmucinous adenocarcinoma.

Article Snippet: The primary antibody against POFUT1 (14929‐1‐AP; Proteintech), PLAGL2 (11540‐1‐ AP; Proteintech), MUC2 (27675‐1‐AP; Proteintech), and β‐actin (ab8227; Abcam) were diluted 1:4000, 1:2500, and 1:1000 respectively, and applied overnight at 4°C.

Techniques: Expressing, Comparison, Immunohistochemistry

Journal: International journal of oncology

Article Title: Protein O-fucosyltransferase 1: a potential diagnostic marker and therapeutic target for human oral cancer.

doi: 10.3892/ijo.2013.2110

Figure Lengend Snippet: Figure 1. Evaluation of POFUT1 expression in OSCC-derived cell lines. (A) POFUT1 mRNA levels are analyzed in OSCC-derived cells and HNOKs by qRT-PCR. Significant upregulation of POFUT1 mRNA is seen in five OSCC-derived cell lines compared with HNOKs (*P<0.05, Mann- Whitney U test). Data are expressed as the means ± SEM in triplicate. (B) Representative western blot data of POFUT1 in OSCC-derived cell lines and HNOKs. Densitometric POFUT1 protein data are normalized to GAPDH protein levels. The values are expressed as a percentage of the HNOKs. POFUT1 protein is upregulated in OSCC-derived cell lines com pared with HNOKs.

Article Snippet: The sections were blocked for 2 h at room temperature with 1.5% blocking serum (Santa Cruz Biotechnology) in PBS before reacting with anti-POFUT1 antibody (1:1,000 dilution) at 4 ̊C in a moist chamber overnight.

Techniques: Expressing, Derivative Assay, Quantitative RT-PCR, MANN-WHITNEY, Western Blot

Journal: International journal of oncology

Article Title: Protein O-fucosyltransferase 1: a potential diagnostic marker and therapeutic target for human oral cancer.

doi: 10.3892/ijo.2013.2110

Figure Lengend Snippet: Figure 2. Evaluation of POFUT1 protein expression in primary OSCCs. (A) Representative IHC results of POFUT1 in normal oral tissue and primary OSCC. (a) Normal oral tissues exhibit negative POFUT1 protein expression. Skeletal muscle tissues are immunostained for POFUT1 (positive control). Original magnification, x100. Scale bars, 50 µm. (b) Positive immunoreactivity for POFUT1 in OSCCs is detected in the cytoplasm. Original magnification, x100. Scale bars, 50 µm. (B) State of POFUT1 protein expression in normal oral tissues (n=128) and primary OSCCs (n=128). The POFUT1 IHC scores are calculated as follows: IHC score = 1 x (number of weakly stained cells in the field) + 2 x (number of moderately stained cells in the field) + 3 x (number of intensely stained cells in the field). The POFUT1 IHC scores for normal oral tissues and OSCCs range from 76.2 to 180.0 (median, 165.8) and 112.5 to 225.5 (median, 192.4), respectively. The POFUT1 protein expression level in OSCCs is significantly (P<0.05, Mann-Whitney U test) higher than that in normal oral tissues. (C) The POFUT1 IHC scores for T3/T4 (149.8 to 240; median, 204.5) are significantly (P<0.05, Mann-Whitney U test) higher than those of T1/T2 (77.8 to 215; median, 189.5).

Article Snippet: The sections were blocked for 2 h at room temperature with 1.5% blocking serum (Santa Cruz Biotechnology) in PBS before reacting with anti-POFUT1 antibody (1:1,000 dilution) at 4 ̊C in a moist chamber overnight.

Techniques: Expressing, Positive Control, Staining, MANN-WHITNEY

Journal: International journal of oncology

Article Title: Protein O-fucosyltransferase 1: a potential diagnostic marker and therapeutic target for human oral cancer.

doi: 10.3892/ijo.2013.2110

Figure Lengend Snippet: Figure 3. Expression of POFUT1 in shPOFUT1-transfected cells. (A) qRT-PCR shows that POFUT1 mRNA expression in the shPOFUT1-transfected cells (KOSC2- and HSC-2-derived tranfectant cells; two clones each) is significantly (*P<0.05, Mann-Whitney U test) lower than that in the mock-transfected cells. (B) Western blot analysis shows that the POFUT1 protein levels in the shPOFUT1-transfected cells (KOSC2- and HSC-2-derived transfectant cells; two clones each) also have decreased markedly compared with that in the mock-transfected cells.

Article Snippet: The sections were blocked for 2 h at room temperature with 1.5% blocking serum (Santa Cruz Biotechnology) in PBS before reacting with anti-POFUT1 antibody (1:1,000 dilution) at 4 ̊C in a moist chamber overnight.

Techniques: Expressing, Transfection, Quantitative RT-PCR, Derivative Assay, Clone Assay, MANN-WHITNEY, Western Blot

Journal: International journal of oncology

Article Title: Protein O-fucosyltransferase 1: a potential diagnostic marker and therapeutic target for human oral cancer.

doi: 10.3892/ijo.2013.2110

Figure Lengend Snippet: Figure 1. Evaluation of POFUT1 expression in OSCC-derived cell lines. (A) POFUT1 mRNA levels are analyzed in OSCC-derived cells and HNOKs by qRT-PCR. Significant upregulation of POFUT1 mRNA is seen in five OSCC-derived cell lines compared with HNOKs (*P<0.05, Mann- Whitney U test). Data are expressed as the means ± SEM in triplicate. (B) Representative western blot data of POFUT1 in OSCC-derived cell lines and HNOKs. Densitometric POFUT1 protein data are normalized to GAPDH protein levels. The values are expressed as a percentage of the HNOKs. POFUT1 protein is upregulated in OSCC-derived cell lines com pared with HNOKs.

Article Snippet: IHC was performed on 4-μm sections of paraffinembedded specimens using rabbit anti-POFUT1 polyclonal antibody (OriGene Technologies).

Techniques: Expressing, Derivative Assay, Quantitative RT-PCR, MANN-WHITNEY, Western Blot

Journal: International journal of oncology

Article Title: Protein O-fucosyltransferase 1: a potential diagnostic marker and therapeutic target for human oral cancer.

doi: 10.3892/ijo.2013.2110

Figure Lengend Snippet: Figure 2. Evaluation of POFUT1 protein expression in primary OSCCs. (A) Representative IHC results of POFUT1 in normal oral tissue and primary OSCC. (a) Normal oral tissues exhibit negative POFUT1 protein expression. Skeletal muscle tissues are immunostained for POFUT1 (positive control). Original magnification, x100. Scale bars, 50 µm. (b) Positive immunoreactivity for POFUT1 in OSCCs is detected in the cytoplasm. Original magnification, x100. Scale bars, 50 µm. (B) State of POFUT1 protein expression in normal oral tissues (n=128) and primary OSCCs (n=128). The POFUT1 IHC scores are calculated as follows: IHC score = 1 x (number of weakly stained cells in the field) + 2 x (number of moderately stained cells in the field) + 3 x (number of intensely stained cells in the field). The POFUT1 IHC scores for normal oral tissues and OSCCs range from 76.2 to 180.0 (median, 165.8) and 112.5 to 225.5 (median, 192.4), respectively. The POFUT1 protein expression level in OSCCs is significantly (P<0.05, Mann-Whitney U test) higher than that in normal oral tissues. (C) The POFUT1 IHC scores for T3/T4 (149.8 to 240; median, 204.5) are significantly (P<0.05, Mann-Whitney U test) higher than those of T1/T2 (77.8 to 215; median, 189.5).

Article Snippet: IHC was performed on 4-μm sections of paraffinembedded specimens using rabbit anti-POFUT1 polyclonal antibody (OriGene Technologies).

Techniques: Expressing, Positive Control, Staining, MANN-WHITNEY

Journal: International journal of oncology

Article Title: Protein O-fucosyltransferase 1: a potential diagnostic marker and therapeutic target for human oral cancer.

doi: 10.3892/ijo.2013.2110

Figure Lengend Snippet: Figure 3. Expression of POFUT1 in shPOFUT1-transfected cells. (A) qRT-PCR shows that POFUT1 mRNA expression in the shPOFUT1-transfected cells (KOSC2- and HSC-2-derived tranfectant cells; two clones each) is significantly (*P<0.05, Mann-Whitney U test) lower than that in the mock-transfected cells. (B) Western blot analysis shows that the POFUT1 protein levels in the shPOFUT1-transfected cells (KOSC2- and HSC-2-derived transfectant cells; two clones each) also have decreased markedly compared with that in the mock-transfected cells.

Article Snippet: IHC was performed on 4-μm sections of paraffinembedded specimens using rabbit anti-POFUT1 polyclonal antibody (OriGene Technologies).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Derivative Assay, Clone Assay, MANN-WHITNEY, Western Blot

Journal: Cell biochemistry and function

Article Title: POFUT1 and PLAGL2 are characteristic markers of mucinous colorectal cancer associated with MUC2 expression.

doi: 10.1002/cbf.3989

Figure Lengend Snippet: FIGURE 4 Effect of POFUT1 and PLAGL2 knockdown on MUC2 expression in WiDr cells. (A) Comparison of POFUT1 and PLAGL2 expression by qRT‐PCR between control and knockdown WiDr cells (n = 3). (B) Comparison of MUC2 and KRT20 expression by qRT‐PCR between control and knockdown WiDr cells (n = 3). (C) Western blot analysis comparing the protein expression of POFUT1, PLAGL2, and MUC2 between control and knocked‐down WiDr cells. β‐actin was analyzed as a loading control. **p < .01; *p < .05.

Article Snippet: The primary antibody against POFUT1 (14929‐1‐AP; Proteintech), PLAGL2 (11540‐1‐ AP; Proteintech), MUC2 (27675‐1‐AP; Proteintech), and β‐actin (ab8227; Abcam) were diluted 1:4000, 1:2500, and 1:1000 respectively, and applied overnight at 4°C.

Techniques: Knockdown, Expressing, Comparison, Quantitative RT-PCR, Control, Western Blot

Journal: Cell biochemistry and function

Article Title: POFUT1 and PLAGL2 are characteristic markers of mucinous colorectal cancer associated with MUC2 expression.

doi: 10.1002/cbf.3989

Figure Lengend Snippet: FIGURE 5 Effect of POFUT1 and PLAGL2 knockdown in vivo. (A) Images of the tumors generated from control and knocked down WiDr cells. Scale bar, 5 mm. (B) Comparison of tumor volumes between control and knocked‐down WiDr‐derived tumors (n = 3). (C) Comparison of MUC2 expression by qRT‐PCR between control and knocked down WiDr‐derived tumors (n = 3). (D) Representative images of IHC on MUC2 in control and knocked down WiDr‐derived tumors. Scale bar, 250 μm. *p < .05.

Article Snippet: The primary antibody against POFUT1 (14929‐1‐AP; Proteintech), PLAGL2 (11540‐1‐ AP; Proteintech), MUC2 (27675‐1‐AP; Proteintech), and β‐actin (ab8227; Abcam) were diluted 1:4000, 1:2500, and 1:1000 respectively, and applied overnight at 4°C.

Techniques: Knockdown, In Vivo, Generated, Control, Comparison, Derivative Assay, Expressing, Quantitative RT-PCR

Journal: Cell biochemistry and function

Article Title: POFUT1 and PLAGL2 are characteristic markers of mucinous colorectal cancer associated with MUC2 expression.

doi: 10.1002/cbf.3989

Figure Lengend Snippet: FIGURE 6 Expression of POFUT1 and PLAGL2 in human CRCs. (A) Representative images of IHC on POFUT1, PLAGL2, and MUC2 in human MAC and NMAC tissues. Scale bar, 200 μm (B) Comparison of IHC scores between MAC and NMAC tissues. (n = 15 each). **p < .01; *p < .05. CRC, colorectal cancer; IHC, immunohistochemistry; MAC, mucinous adenocarcinoma; NMAC, nonmucinous adenocarcinoma.

Article Snippet: The primary antibody against POFUT1 (14929‐1‐AP; Proteintech), PLAGL2 (11540‐1‐ AP; Proteintech), MUC2 (27675‐1‐AP; Proteintech), and β‐actin (ab8227; Abcam) were diluted 1:4000, 1:2500, and 1:1000 respectively, and applied overnight at 4°C.

Techniques: Expressing, Comparison, Immunohistochemistry

Journal: EMBO Molecular Medicine

Article Title: A spoonful of L‐fucose—an efficient therapy for GFUS‐CDG, a new glycosylation disorder

doi: 10.15252/emmm.202114332

Figure Lengend Snippet: Western blot analysis of FUT8, POFUT1 and POFUT2. Expression of the proteins was analysed by western blotting on a 10% SDS–PAGE with cytosolic fractions derived from control and patient fibroblasts. Data were obtained from fibroblasts; n = 3, for statistics an unpaired t ‐test was performed. qPCR studies on FUT8, POFUT1 and POFUT2 . mRNA expression revealed a significantly increased transcript level of FUT8 (2.79 fold change; ± 1,10 fold change; * P = 0.0498) normalized to a control. Expression of POFUT1 and POFUT2 was not changed significantly. Data were obtained from fibroblasts; n = 9; experiment was independently repeated three times, for statistics an ANOVA was performed. Expression of ADAMTS13 in sera. The amount of ADAMTS13 secreted into sera of controls and the patient before and after 8 weeks of L‐fucose treatment was detected by western blot and normalized to transferrin. Data were obtained from serum; n = 12 for controls. Data information: * P < 0.05; ** P < 0.01. Bars and error bars represent mean ± SD. Exact P ‐values are reported in the results part.

Article Snippet: For FUT8 (Proteintech, 66118‐1‐lg, mouse anti‐human), POFUT1 (Proteintech, 14929‐1‐AP, rabbit anti‐human), POFUT2 (Proteintech, 17764‐1‐AP, rabbit anti‐human) and ADAMTS13 (Thermo Fisher, MA5‐34796, rabbit anti‐human), the procedure was performed analogously.

Techniques: Western Blot, Expressing, SDS Page, Derivative Assay, Control

Journal: Cell Proliferation

Article Title: Epiregulin promotes trophoblast epithelial–mesenchymal transition through poFUT1 and O‐fucosylation by poFUT1 on uPA

doi: 10.1111/cpr.12745

Figure Lengend Snippet: Epiregulin promotes EMT of human trophoblast cells. A, JAR and (B) HTR‐8/SVneo cells were treated with epiregulin at different concentrations (0, 10, 50, 100 ng/mL) and with 50 ng/mL epiregulin for different times (0, 24, 48, 72 h). Total RNA was analysed for E‐cadherin and N‐cadherin expression by RT‐PCR, with GAPDH serving as the internal control. C, JAR and (D) HTR‐8/SVneo cell lysates were prepared for immunoblotting analysis for E‐cadherin, N‐cadherin and vimentin, and GAPDH served as the internal control. E and F, Analysis of MMP9 activity by gelatine zymography. G and H, Cell extension, and invasive ability were examined by Transwell invasion assays. I, JAR and (J) HTR‐8/SVneo cells were analysed for E‐cadherin (green) and N‐cadherin (red) by immunofluorescent staining. DAPI was used for nuclear staining. Scale bars: 50 μm. The statistical analysis was shown: * P < .05; ** P < .01; *** P < .001. EMT, epithelial–mesenchymal transition

Article Snippet: The membranes were incubated at 4°C overnight with the primary antibodies: uPA and uPAR (Abcam), p‐c‐FOS (Ser 32 ), p‐c‐JUN (Ser 73 ), c‐FOS, c‐JUN, Akt, p‐Akt (Tyr 308 ), PDK and p‐PDK (Ser 241 ) (Cell Signaling Technology) and CK‐7, vimentin, N‐cadherin, E‐cadherin, poFUT1, HLA‐G and GAPDH (Proteintech).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Western Blot, Activity Assay, Zymography, Staining

Journal: EBioMedicine

Article Title: poFUT1 promotes endometrial decidualization by enhancing the O-fucosylation of Notch1

doi: 10.1016/j.ebiom.2019.05.027

Figure Lengend Snippet: poFUT1 increases hESCs proliferation ability. hESCs were transfected with vector, poFUT1 cDNA or scramble RNA, or poFUT1 siRNA. (A) CCK-8 assay analysis was used to evaluate the cell proliferation capabilities of hESCs. (B, C) Western blot analysis of the expression of PCNA, cyclin D1, CDK2, CDK6 and GAPDH in hESCs. (D, E) Representative immnofluorescent staining of PCNA in hESCs. (bar:50 μm).

Article Snippet: Primary antibodies: poFUT1 (1:100, Proteintech, Wuhan, China), IGFBP1 (1:100, Santa Cruz, USA) and PCNA (1:100, Proteintech, Wuhan, China) were applied at 4 °C overnight in a wet chamber.

Techniques: Transfection, Plasmid Preparation, CCK-8 Assay, Western Blot, Expressing, Staining

Journal: EBioMedicine

Article Title: poFUT1 promotes endometrial decidualization by enhancing the O-fucosylation of Notch1

doi: 10.1016/j.ebiom.2019.05.027

Figure Lengend Snippet: poFUT1 siRNA inhibits decidualization in vivo. The murine uterus was injected with normal saline (NS) or poFUT1 siRNA on Day 3 of pregnancy. (A) The number of implanted embryos in the uterus on Day 8 of pregnancy. (B) Immunohistochemical analysis of poFUT1, IGFBP1 and PCNA in the uterine endometrium of pregnant mice. (bar: 200 μm).

Article Snippet: Primary antibodies: poFUT1 (1:100, Proteintech, Wuhan, China), IGFBP1 (1:100, Santa Cruz, USA) and PCNA (1:100, Proteintech, Wuhan, China) were applied at 4 °C overnight in a wet chamber.

Techniques: In Vivo, Injection, Saline, Immunohistochemical staining