Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: The glycogene alterations and potential effects in esophageal squamous cell carcinoma

doi: 10.1007/s00018-024-05534-3

Figure Lengend Snippet: POFUT1 is highly expressed and is a poor prognostic factor in ESCC. A POFUT1 expression in ESCC and normal tissues from Liu’s published proteome, our unpublished proteome data and previously published transcriptome data. B Representative images and quantification of POFUT1 expression in the cytoplasm and nucleus of ESCC tissues and paired normal tissues (n = 85) detected by IHC. Scale bar, 100 μm. C Kaplan–Meier plot presenting the correlation between high POFUT1 expression and poor prognosis in ESCC patients and in male ESCC patients, patients with a history of alcohol consumption, and patients with stage T3. D Univariate and multifactorial Cox regression analyses of the association of POFUT1 levels with clinicopathological factors in ESCC patients

Article Snippet: After being blocked, the slides were incubated with POFUT1 antibody (Proteintech) overnight.

Techniques: Expressing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: The glycogene alterations and potential effects in esophageal squamous cell carcinoma

doi: 10.1007/s00018-024-05534-3

Figure Lengend Snippet: Knockdown of POFUT1 suppresses the proliferation, metastasis and invasion of ESCC. A POFUT1 protein expression was examined in 6 kinds of cells by Western Blot. B , C RT-qPCR ( B ) and Western Blot ( C ) revealed that POFUT1 was efficiently knocked down in KYSE150 and TE-1 cells. D , E Analysis of the proliferation ability of POFUT1 knockdown KYSE150 and TE-1 cells by MTT ( D ) and colony formation assays ( E ). F Analysis of the migration and invasion abilities of POFUT1-knockdown KYSE150 and TE-1 cells by transwell assays. G Analysis of E-cadherin, N-cadherin and vimentin expression in POFUT1-knockdown KYSE150 and TE-1 cells by Western Blot. Scale bar, 200 μm. **, P < 0.01; ***, P < 0.001

Article Snippet: After being blocked, the slides were incubated with POFUT1 antibody (Proteintech) overnight.

Techniques: Knockdown, Expressing, Western Blot, Quantitative RT-PCR, Migration

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: The glycogene alterations and potential effects in esophageal squamous cell carcinoma

doi: 10.1007/s00018-024-05534-3

Figure Lengend Snippet: POFUT1 overexpression promotes the proliferation, metastasis and invasion of ESCC. A , B RT-qPCR ( A ) and Western Blot ( B ) revealed that POFUT1 was efficiently overexpressed in KYSE450, TE-1 and KYSE150 cells. C , D Analysis of the proliferation of POFUT1-overexpression KYSE450 cells by MTT ( C ) and colony formation assays ( D ). E Analysis of the migration and invasion abilities of POFUT1-overexpression KYSE450, TE-1 and KYSE150 cells by transwell assays. F Analysis of E-cadherin, N-cadherin and vimentin expression in POFUT1-overexpression KYSE450 and TE-1 cells by Western Blot. G Quantitation of the number of lung metastatic nodules in mice in the NC group and POFUT1-OE group. H Representative images and HE staining of lung tissues from mice in the NC group and POFUT1-OE group. Scale bar, 200 μm. *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: After being blocked, the slides were incubated with POFUT1 antibody (Proteintech) overnight.

Techniques: Over Expression, Quantitative RT-PCR, Western Blot, Migration, Expressing, Quantitation Assay, Staining

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: The glycogene alterations and potential effects in esophageal squamous cell carcinoma

doi: 10.1007/s00018-024-05534-3

Figure Lengend Snippet: The overexpression of POFUT1 increases the overall O-fucosylation level and activates the Notch signaling pathway, which mediates POFUT1 induced pro-migration in ESCC. A Analysis of the level of overall cellular O-fucosylation modification after POFUT1 knockdown in KYSE150 and TE-1 cells by click chemistry reaction. B Analysis of the level of overall O-fucosylation modification after POFUT1 overexpression in KYSE450 and TE-1 cells by click chemistry reaction. C Analysis of the protein level and fucosylation modification level of Notch1 in KYSE450 and TE-1 cells stably overexpressing POFUT1 and the control using AAL lectin enrichment followed by Western Blot. D , E Analysis of the protein expression of NICD ( D ) and HES1 ( E ) after POFUT1 was knocked down in KYSE150 and TE-1 cells, and POFUT1 was overexpressed in KYSE450 and TE-1 cells. F Analysis of the migration and invasion abilities of KYSE450 and TE-1 cells stably overexpressing POFUT1, with or without DAPT treatment by transwell assay. **, P < 0.01; ***, P < 0.001

Article Snippet: After being blocked, the slides were incubated with POFUT1 antibody (Proteintech) overnight.

Techniques: Over Expression, Migration, Modification, Knockdown, Stable Transfection, Control, Western Blot, Expressing, Transwell Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: The glycogene alterations and potential effects in esophageal squamous cell carcinoma

doi: 10.1007/s00018-024-05534-3

Figure Lengend Snippet: POFUT1 is highly expressed and is a poor prognostic factor in ESCC. A POFUT1 expression in ESCC and normal tissues from Liu’s published proteome, our unpublished proteome data and previously published transcriptome data. B Representative images and quantification of POFUT1 expression in the cytoplasm and nucleus of ESCC tissues and paired normal tissues (n = 85) detected by IHC. Scale bar, 100 μm. C Kaplan–Meier plot presenting the correlation between high POFUT1 expression and poor prognosis in ESCC patients and in male ESCC patients, patients with a history of alcohol consumption, and patients with stage T3. D Univariate and multifactorial Cox regression analyses of the association of POFUT1 levels with clinicopathological factors in ESCC patients

Article Snippet: After being blocked, the membrane was incubated with a diluted primary antibody (RPN1, Abcam, 1:1000) (POFUT1, Proteintech, 1:1000) at 4 °C overnight.

Techniques: Expressing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: The glycogene alterations and potential effects in esophageal squamous cell carcinoma

doi: 10.1007/s00018-024-05534-3

Figure Lengend Snippet: Knockdown of POFUT1 suppresses the proliferation, metastasis and invasion of ESCC. A POFUT1 protein expression was examined in 6 kinds of cells by Western Blot. B , C RT-qPCR ( B ) and Western Blot ( C ) revealed that POFUT1 was efficiently knocked down in KYSE150 and TE-1 cells. D , E Analysis of the proliferation ability of POFUT1 knockdown KYSE150 and TE-1 cells by MTT ( D ) and colony formation assays ( E ). F Analysis of the migration and invasion abilities of POFUT1-knockdown KYSE150 and TE-1 cells by transwell assays. G Analysis of E-cadherin, N-cadherin and vimentin expression in POFUT1-knockdown KYSE150 and TE-1 cells by Western Blot. Scale bar, 200 μm. **, P < 0.01; ***, P < 0.001

Article Snippet: After being blocked, the membrane was incubated with a diluted primary antibody (RPN1, Abcam, 1:1000) (POFUT1, Proteintech, 1:1000) at 4 °C overnight.

Techniques: Knockdown, Expressing, Western Blot, Quantitative RT-PCR, Migration

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: The glycogene alterations and potential effects in esophageal squamous cell carcinoma

doi: 10.1007/s00018-024-05534-3

Figure Lengend Snippet: POFUT1 overexpression promotes the proliferation, metastasis and invasion of ESCC. A , B RT-qPCR ( A ) and Western Blot ( B ) revealed that POFUT1 was efficiently overexpressed in KYSE450, TE-1 and KYSE150 cells. C , D Analysis of the proliferation of POFUT1-overexpression KYSE450 cells by MTT ( C ) and colony formation assays ( D ). E Analysis of the migration and invasion abilities of POFUT1-overexpression KYSE450, TE-1 and KYSE150 cells by transwell assays. F Analysis of E-cadherin, N-cadherin and vimentin expression in POFUT1-overexpression KYSE450 and TE-1 cells by Western Blot. G Quantitation of the number of lung metastatic nodules in mice in the NC group and POFUT1-OE group. H Representative images and HE staining of lung tissues from mice in the NC group and POFUT1-OE group. Scale bar, 200 μm. *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: After being blocked, the membrane was incubated with a diluted primary antibody (RPN1, Abcam, 1:1000) (POFUT1, Proteintech, 1:1000) at 4 °C overnight.

Techniques: Over Expression, Quantitative RT-PCR, Western Blot, Migration, Expressing, Quantitation Assay, Staining

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: The glycogene alterations and potential effects in esophageal squamous cell carcinoma

doi: 10.1007/s00018-024-05534-3

Figure Lengend Snippet: The overexpression of POFUT1 increases the overall O-fucosylation level and activates the Notch signaling pathway, which mediates POFUT1 induced pro-migration in ESCC. A Analysis of the level of overall cellular O-fucosylation modification after POFUT1 knockdown in KYSE150 and TE-1 cells by click chemistry reaction. B Analysis of the level of overall O-fucosylation modification after POFUT1 overexpression in KYSE450 and TE-1 cells by click chemistry reaction. C Analysis of the protein level and fucosylation modification level of Notch1 in KYSE450 and TE-1 cells stably overexpressing POFUT1 and the control using AAL lectin enrichment followed by Western Blot. D , E Analysis of the protein expression of NICD ( D ) and HES1 ( E ) after POFUT1 was knocked down in KYSE150 and TE-1 cells, and POFUT1 was overexpressed in KYSE450 and TE-1 cells. F Analysis of the migration and invasion abilities of KYSE450 and TE-1 cells stably overexpressing POFUT1, with or without DAPT treatment by transwell assay. **, P < 0.01; ***, P < 0.001

Article Snippet: After being blocked, the membrane was incubated with a diluted primary antibody (RPN1, Abcam, 1:1000) (POFUT1, Proteintech, 1:1000) at 4 °C overnight.

Techniques: Over Expression, Migration, Modification, Knockdown, Stable Transfection, Control, Western Blot, Expressing, Transwell Assay

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