integrin α6 expression  (Human Protein Atlas)

Journal: Nature Communications

Article Title: Dual Role of WISP1 in maintaining glioma stem cells and tumor-supportive macrophages in glioblastoma

doi: 10.1038/s41467-020-16827-z

Figure Lengend Snippet: a , b , Immunoblot analysis of Akt activating phosphorylation (Ser473) in T4121 GSCs ( a ) and T387 GSCs ( b ) treated with 5 μg/ml Integrin blocking antibody (Anti-Integrin ab) or relative isotype IgG control in combination with WISP1 overexpression or vector control for 48 h. α3, Integrin α3; α6, Integrin α6; α7, Integrin α7. c , d , Cell viability ( c ) or tumorsphere formation ( d ) assay of T4121 GSCs treated with 5 μg/ml Integrin α6 blocking antibody (ab) or isotype IgG in combination with WISP1 overexpression or vector control. n = 6 biological independent samples. Data are shown as means ± s.d. **** p <0.0001, two way ANOVA analysis followed by Tukey’s multiple test ( c ). **** p <0.0001, two-tailed unpaired t -test ( d ). e , f , Cell viability ( e ) or tumorsphere formation ( f ) assay of T4121 GSCs treated with Integrin β1 blocking antibody (5 μg/ml) or isotype IgG in combination with WISP1 overexpression or vector control. n = 5 ( e ) or 6 ( f ) biological independent samples. Data are represented as mean ± s.d. **** p <0.0001, two way ANOVA analysis followed by Tukey’s multiple test ( e ). **** p <0.0001, two-tailed unpaired t -test ( f ). g , h , Cell viability ( g ) or tumorsphere formation ( h ) assay of T4121 GSCs treated with Integrin blocking antibody (5 μg/ml) or isotype IgG for 6 days. n = 5 biological independent samples. Data are represented as mean ± s.d. *** p = 0.001, **** p <0.0001, two-tailed unpaired t -test. α6, Integrin α6; β1, Integrin β1; β4, Integrin β4. i Immunoblot analysis of Akt phosphorylation (Ser473) and Integrin α6 expression in T4121 GSCs transduced with shIntegrin α6 or shNT control. j , k , Cell viability ( j ) or tumorsphere formation ( k ) assay of T4121 GSCs transduced with shIntegrin α6 or shNT. n = 5 biological independent samples. Data are shown as means ± s.d. **** p <0.0001, two way ANOVA analysis followed by Tukey’s multiple test ( j ). **** p <0.0001, two-tailed unpaired t -test ( k ). Source data are provided as a file.

Article Snippet: Lentiviral clones expressing two non-overlapping shRNAs against human WISP1 (TRCN0000373969, TRCN0000373970), human Integrin α6 (TRCN0000296162, TRCN000057775) and non-targeting shRNA (SHC002) were obtained from Sigma-Aldrich.

Techniques: Western Blot, Blocking Assay, Over Expression, Plasmid Preparation, Two Tailed Test, Expressing, Transduction

Journal: Nature Communications

Article Title: Dual Role of WISP1 in maintaining glioma stem cells and tumor-supportive macrophages in glioblastoma

doi: 10.1038/s41467-020-16827-z

Figure Lengend Snippet: a , b , CoIP assays of protein interaction in T4121 GSCs ( a ) or T387 GSCs ( b ) transduced with WISP1 overexpression lentivirus. Cell lysates were immunoprecipitated (IP) with anti-Integrin α6 antibody and then immunoblotted with anti-WISP1, anti-Integrin α6 and anti-Integrin β1 antibodies. c , d , CoIP assays of protein interaction in T4121 GSCs ( c ) or T387 GSCs ( d ) transduced with WISP1 overexpression lentivirus. Cell lysates were immunoprecipitated (IP) with anti-Integrin β1 antibody and then immunoblotted with anti-WISP1, anti-Integrin α6 and anti-Integrin β1 antibodies. e Immunoblot analysis of Akt phosphorylation (Ser473) in T4121 GSCs treated with 5 or 10 μg/ml Integrin blocking antibody in combination with 0.2 or 0.8 μg/ml rWISP1 protein for 12 h. f Immunofluorescent staining of Integrin α6 (green) and WISP1 (red) in human primary GBM samples. Scale bar, 30 μM.

Article Snippet: Lentiviral clones expressing two non-overlapping shRNAs against human WISP1 (TRCN0000373969, TRCN0000373970), human Integrin α6 (TRCN0000296162, TRCN000057775) and non-targeting shRNA (SHC002) were obtained from Sigma-Aldrich.

Techniques: Transduction, Over Expression, Immunoprecipitation, Western Blot, Blocking Assay, Staining

Journal: Nature Communications

Article Title: Dual Role of WISP1 in maintaining glioma stem cells and tumor-supportive macrophages in glioblastoma

doi: 10.1038/s41467-020-16827-z

Figure Lengend Snippet: WISP1 is preferentially expressed and secreted by GSCs and promotes the GSC maintenance through an autocrine loop to activate Integrin α6β1-Akt signaling. GSC-secreted WISP1 also promotes M2 TAM survival through Integrin α6β1-mediated Akt activation in a paracrine manner. The dual role of WISP1 augments GBM malignant growth and progression. Targeting Wnt/β-catenin-WISP1 signaling with carnosic acid (CA) potently inhibits GBM tumor growth and may have therapeutic potential. Reprinted with permission, Cleveland Clinic Center for Medical Art & Photography © 2019–2020. All Rights Reserved.

Article Snippet: Lentiviral clones expressing two non-overlapping shRNAs against human WISP1 (TRCN0000373969, TRCN0000373970), human Integrin α6 (TRCN0000296162, TRCN000057775) and non-targeting shRNA (SHC002) were obtained from Sigma-Aldrich.

Techniques: Activation Assay