rabbit polyclonal antibody against pd l1  (Novus Biologicals)

Journal: Cells

Article Title: The Immune Checkpoint Protein PD-L1 Regulates Ciliogenesis and Hedgehog Signaling

doi: 10.3390/cells13121003

Figure Lengend Snippet: PD-L1 is located at the centrosome and Golgi. ( A ) Western blot analysis indicating the differential expression of PD-L1 in fibroblast and epithelial cells. ( B ) NIH3T3 cells stained with PD-L1 (green) antibody and co-stained with centrosome marker, γ-tubulin (red) (top panel), and cilium marker, acetylated-α-tubulin (red) (bottom panel). Centrosome localized PD-L1 is indicated with the white arrow. ( C ) RCTE cells stained with PD-L1 (green) antibody and co-stained with γ-tubulin (red) (top panel) and acetylated-α-tubulin (red) (bottom panel). Cells were serum starved to induce cilia growth. ( D ) RCTE cells stained with PD-L1 (green) antibody and co-stained with Golgi marker, GIANTIN (red). ( E ) Overexpression of GFP-PD-L1 (green) co-stained with Golgi marker, Giantin (red) in RCTE cells. ( F ) Overexpression of GFP-PD-L1 (green) co-stained with Golgi-associated protein Ift20 (red) in NIH3T3 cells. All cells were counterstained with DAPI (blue). Scale bars, 20 μm.

Article Snippet: Primary antibodies used in this study are listed as follows: mouse monoclonal antibodies against acetylated α-tubulin (6-11B-1, Sigma, St. Louis, MO, USA, T7451, 1:4000 used for immunofluorescence [IF]), γ-tubulin (GTU-88, Sigma, T5326, 1:1000 used for IF), α-tubulin (DM1A, sc-32293, 1:1000), Ninein (F-7, Santa Cruz, Dallas, TX, USA, sc-390540, 1:300 for IF), Cep164 (E-9, Santa Cruz, sc-515403, 1:300 used for IF), C-nap1 (E-9, Santa Cruz, sc-515403, 1:300 used for IF), Smo (E-5, Santa Cruz, sc-166685, 1:100 used for IF), Gli1 (C-1, Santa Cruz, sc-515751, 1:500 used for WB), Giantin (9B6, Abcam, Waltham, MA, USA, ab37266, 1:500 used for IF), actin (AC-15, Sigma, A1978, 1:3000 used for Western blot [WB]), GFP-tag (B-2, sc-9996, 1:1000 for WB), and Myc (9E10, sc-40, 1:1000 for WB); rabbit polyclonal antibodies against PD-L1 (Proteintech, Rosemont, IL, USA, 17952-1-AP, 1:300 used for IF, and 1:1000 used for WB), Cep135 (Abcam, ab75005, 1:300 used for IF), Ift140 (Proteintech, 17460-1-AP, 1:300 used for IF, and 1:1000 for WB), Ift20 (Proteintech, 13615-1-AP, 1:300 used for IF, and 1:1000 for WB), Rab8a (Proteintech, 55296-1-AP, 1:300 used for IF, and 1:1000 for WB), BBS5 (Proteintech, 14569-1-AP, 1:300 used for IF, and 1:1000 for WB), ARL13B (Proteintech, 17711-1-AP, 1:2000 for IF), GFP-tag (Proteintech, 50430-2-AP, 1:1000 for WB) and Myc-Tag (CST, 71D10, 1:1000 used for WB); goat polyclonal antibody against polycystin 2 (E-20, Santa Cruz, sc-10377, 1:500 for WB).

Techniques: Western Blot, Quantitative Proteomics, Staining, Marker, Over Expression

Journal: Cells

Article Title: The Immune Checkpoint Protein PD-L1 Regulates Ciliogenesis and Hedgehog Signaling

doi: 10.3390/cells13121003

Figure Lengend Snippet: PD-L1 regulates ciliogenesis in 3T3 cells. ( A ) Representative images of acetylated-α-tubulin (red) co-stained with Arl13B (green) in PD-L1 siRNA knockdown NIH3T3 cells compared to control siRNA cells. ( B ) Quantitative data of cilium length ( n > 100) in PD-L1 siRNA knockdown NIH3T3 cells compared to control siRNA cells. ( C – E ) Representative images of acetylated-α-tubulin (red) and GFP-PD-L1 (green) ( C ), and quantitative data of percentage ciliated cells ( n > 75) ( D ), and cilium length ( n > 75) ( E ), in NIH3T3 cells transfected with GFP-PD-L1. All cells were counterstained with DAPI (blue). The quantitative data was obtained by analyzing only cells that were GFP-positive. Scale bar, 20 μm.

Article Snippet: Primary antibodies used in this study are listed as follows: mouse monoclonal antibodies against acetylated α-tubulin (6-11B-1, Sigma, St. Louis, MO, USA, T7451, 1:4000 used for immunofluorescence [IF]), γ-tubulin (GTU-88, Sigma, T5326, 1:1000 used for IF), α-tubulin (DM1A, sc-32293, 1:1000), Ninein (F-7, Santa Cruz, Dallas, TX, USA, sc-390540, 1:300 for IF), Cep164 (E-9, Santa Cruz, sc-515403, 1:300 used for IF), C-nap1 (E-9, Santa Cruz, sc-515403, 1:300 used for IF), Smo (E-5, Santa Cruz, sc-166685, 1:100 used for IF), Gli1 (C-1, Santa Cruz, sc-515751, 1:500 used for WB), Giantin (9B6, Abcam, Waltham, MA, USA, ab37266, 1:500 used for IF), actin (AC-15, Sigma, A1978, 1:3000 used for Western blot [WB]), GFP-tag (B-2, sc-9996, 1:1000 for WB), and Myc (9E10, sc-40, 1:1000 for WB); rabbit polyclonal antibodies against PD-L1 (Proteintech, Rosemont, IL, USA, 17952-1-AP, 1:300 used for IF, and 1:1000 used for WB), Cep135 (Abcam, ab75005, 1:300 used for IF), Ift140 (Proteintech, 17460-1-AP, 1:300 used for IF, and 1:1000 for WB), Ift20 (Proteintech, 13615-1-AP, 1:300 used for IF, and 1:1000 for WB), Rab8a (Proteintech, 55296-1-AP, 1:300 used for IF, and 1:1000 for WB), BBS5 (Proteintech, 14569-1-AP, 1:300 used for IF, and 1:1000 for WB), ARL13B (Proteintech, 17711-1-AP, 1:2000 for IF), GFP-tag (Proteintech, 50430-2-AP, 1:1000 for WB) and Myc-Tag (CST, 71D10, 1:1000 used for WB); goat polyclonal antibody against polycystin 2 (E-20, Santa Cruz, sc-10377, 1:500 for WB).

Techniques: Staining, Knockdown, Control, Transfection

Journal: Cells

Article Title: The Immune Checkpoint Protein PD-L1 Regulates Ciliogenesis and Hedgehog Signaling

doi: 10.3390/cells13121003

Figure Lengend Snippet: PD-L1 affects the Golgi accumulation of Ift20 and ciliary protein trafficking of Rab8a and BBS5. ( A ) NIH3T3 cells co-stained for acetylated-α-tubulin and Ift20 (green) in PD-L1 siRNA knockdown NIH3T3 cells compared to control siRNA cells. ( B ) Quantitative data of fluorescence intensity of Ift20 in PD-L1 siRNA knockdown NIH3T3 cells compared to control siRNA cells ( n > 75). ( C ) NIH3T3 cells co-stained for acetylated-α-tubulin and the small GTPase Rab8a (green), in PD-L1 siRNA knockdown NIH3T3 cells compared to control siRNA cells. ( D ) Quantitative data of Rab8a-positive cilia in PD-L1 siRNA knockdown NIH3T3 cells compared to control siRNA cells ( n > 75). ( E ) NIH3T3 cells co-stained for acetylated-α-tubulin and BBS5 (green), in PD-L1 siRNA knockdown NIH3T3 cells compared to control siRNA cells. ( F ) Quantitative data of BBS5-positive cilia in PD-L1 siRNA knockdown NIH3T3 cells compared to control siRNA cells ( n > 60). All cells were counterstained with DAPI (blue). Scale bar, 20 μm.

Article Snippet: Primary antibodies used in this study are listed as follows: mouse monoclonal antibodies against acetylated α-tubulin (6-11B-1, Sigma, St. Louis, MO, USA, T7451, 1:4000 used for immunofluorescence [IF]), γ-tubulin (GTU-88, Sigma, T5326, 1:1000 used for IF), α-tubulin (DM1A, sc-32293, 1:1000), Ninein (F-7, Santa Cruz, Dallas, TX, USA, sc-390540, 1:300 for IF), Cep164 (E-9, Santa Cruz, sc-515403, 1:300 used for IF), C-nap1 (E-9, Santa Cruz, sc-515403, 1:300 used for IF), Smo (E-5, Santa Cruz, sc-166685, 1:100 used for IF), Gli1 (C-1, Santa Cruz, sc-515751, 1:500 used for WB), Giantin (9B6, Abcam, Waltham, MA, USA, ab37266, 1:500 used for IF), actin (AC-15, Sigma, A1978, 1:3000 used for Western blot [WB]), GFP-tag (B-2, sc-9996, 1:1000 for WB), and Myc (9E10, sc-40, 1:1000 for WB); rabbit polyclonal antibodies against PD-L1 (Proteintech, Rosemont, IL, USA, 17952-1-AP, 1:300 used for IF, and 1:1000 used for WB), Cep135 (Abcam, ab75005, 1:300 used for IF), Ift140 (Proteintech, 17460-1-AP, 1:300 used for IF, and 1:1000 for WB), Ift20 (Proteintech, 13615-1-AP, 1:300 used for IF, and 1:1000 for WB), Rab8a (Proteintech, 55296-1-AP, 1:300 used for IF, and 1:1000 for WB), BBS5 (Proteintech, 14569-1-AP, 1:300 used for IF, and 1:1000 for WB), ARL13B (Proteintech, 17711-1-AP, 1:2000 for IF), GFP-tag (Proteintech, 50430-2-AP, 1:1000 for WB) and Myc-Tag (CST, 71D10, 1:1000 used for WB); goat polyclonal antibody against polycystin 2 (E-20, Santa Cruz, sc-10377, 1:500 for WB).

Techniques: Staining, Knockdown, Control, Fluorescence

Journal: Cells

Article Title: The Immune Checkpoint Protein PD-L1 Regulates Ciliogenesis and Hedgehog Signaling

doi: 10.3390/cells13121003

Figure Lengend Snippet: PD-L1 affects the ciliary recruitment of PC-2 and Ift140. ( A ) NIH3T3 cells co-stained for acetylated-α-tubulin and the PC-2 (polycystin 2) (green), in PD-L1 siRNA knockdown NIH3T3 cells compared to control siRNA cells. ( B ) Quantitative data of PC-2-positive cilia in PD-L1 siRNA knockdown NIH3T3 cells compared to control siRNA cells ( n > 80). ( C ) NIH3T3 cells co-stained for acetylated-α-tubulin and the Ift140 (green), in PD-L1 siRNA knockdown NIH3T3 cells compared to control siRNA cells. ( D ) Quantitative data of Ift140-positive cilia in PD-L1 siRNA knockdown NIH3T3 cells compared to control siRNA cells ( n > 80). All cells were counterstained with DAPI (blue). Scale bars, 20 μm.

Article Snippet: Primary antibodies used in this study are listed as follows: mouse monoclonal antibodies against acetylated α-tubulin (6-11B-1, Sigma, St. Louis, MO, USA, T7451, 1:4000 used for immunofluorescence [IF]), γ-tubulin (GTU-88, Sigma, T5326, 1:1000 used for IF), α-tubulin (DM1A, sc-32293, 1:1000), Ninein (F-7, Santa Cruz, Dallas, TX, USA, sc-390540, 1:300 for IF), Cep164 (E-9, Santa Cruz, sc-515403, 1:300 used for IF), C-nap1 (E-9, Santa Cruz, sc-515403, 1:300 used for IF), Smo (E-5, Santa Cruz, sc-166685, 1:100 used for IF), Gli1 (C-1, Santa Cruz, sc-515751, 1:500 used for WB), Giantin (9B6, Abcam, Waltham, MA, USA, ab37266, 1:500 used for IF), actin (AC-15, Sigma, A1978, 1:3000 used for Western blot [WB]), GFP-tag (B-2, sc-9996, 1:1000 for WB), and Myc (9E10, sc-40, 1:1000 for WB); rabbit polyclonal antibodies against PD-L1 (Proteintech, Rosemont, IL, USA, 17952-1-AP, 1:300 used for IF, and 1:1000 used for WB), Cep135 (Abcam, ab75005, 1:300 used for IF), Ift140 (Proteintech, 17460-1-AP, 1:300 used for IF, and 1:1000 for WB), Ift20 (Proteintech, 13615-1-AP, 1:300 used for IF, and 1:1000 for WB), Rab8a (Proteintech, 55296-1-AP, 1:300 used for IF, and 1:1000 for WB), BBS5 (Proteintech, 14569-1-AP, 1:300 used for IF, and 1:1000 for WB), ARL13B (Proteintech, 17711-1-AP, 1:2000 for IF), GFP-tag (Proteintech, 50430-2-AP, 1:1000 for WB) and Myc-Tag (CST, 71D10, 1:1000 used for WB); goat polyclonal antibody against polycystin 2 (E-20, Santa Cruz, sc-10377, 1:500 for WB).

Techniques: Staining, Knockdown, Control

Journal: Cells

Article Title: The Immune Checkpoint Protein PD-L1 Regulates Ciliogenesis and Hedgehog Signaling

doi: 10.3390/cells13121003

Figure Lengend Snippet: PD-L1 regulates PC-2 cilia localization in a BBS5-independent manner. ( A ) Western blot analysis evaluating the protein level of BBS5 after siRNA knockdown in NIH3T3 cells compared to control siRNA cells. ( B ) Representative images of acetylated-α-tubulin (red) co-stained with Arl13B (green) in BBS5 siRNA knockdown NIH3T3 cells compared to control siRNA cells. ( C ) Quantitative data of cilium length in BBS5 siRNA knockdown NIH3T3 cells compared to control siRNA cells ( n > 100). ( D ) Representative images of acetylated-α-tubulin (red) co-stained with PC-2 (green) in PD-L1 and BBS5 single knockdown, and PD-L1:BBS5 siRNA double knockdown in 3T3 cells compared to control siRNA cells. ( E ) Quantitative data of PC-2-positive cilia in PD-L1 and BBS5 single knockdown, and PD-L1:BBS5 siRNA double knockdown in 3T3 cells compared to control siRNA cells ( n > 75). All cells were counterstained with DAPI (blue). “ns” implies not significant. Scale bars, 20 μm.

Article Snippet: Primary antibodies used in this study are listed as follows: mouse monoclonal antibodies against acetylated α-tubulin (6-11B-1, Sigma, St. Louis, MO, USA, T7451, 1:4000 used for immunofluorescence [IF]), γ-tubulin (GTU-88, Sigma, T5326, 1:1000 used for IF), α-tubulin (DM1A, sc-32293, 1:1000), Ninein (F-7, Santa Cruz, Dallas, TX, USA, sc-390540, 1:300 for IF), Cep164 (E-9, Santa Cruz, sc-515403, 1:300 used for IF), C-nap1 (E-9, Santa Cruz, sc-515403, 1:300 used for IF), Smo (E-5, Santa Cruz, sc-166685, 1:100 used for IF), Gli1 (C-1, Santa Cruz, sc-515751, 1:500 used for WB), Giantin (9B6, Abcam, Waltham, MA, USA, ab37266, 1:500 used for IF), actin (AC-15, Sigma, A1978, 1:3000 used for Western blot [WB]), GFP-tag (B-2, sc-9996, 1:1000 for WB), and Myc (9E10, sc-40, 1:1000 for WB); rabbit polyclonal antibodies against PD-L1 (Proteintech, Rosemont, IL, USA, 17952-1-AP, 1:300 used for IF, and 1:1000 used for WB), Cep135 (Abcam, ab75005, 1:300 used for IF), Ift140 (Proteintech, 17460-1-AP, 1:300 used for IF, and 1:1000 for WB), Ift20 (Proteintech, 13615-1-AP, 1:300 used for IF, and 1:1000 for WB), Rab8a (Proteintech, 55296-1-AP, 1:300 used for IF, and 1:1000 for WB), BBS5 (Proteintech, 14569-1-AP, 1:300 used for IF, and 1:1000 for WB), ARL13B (Proteintech, 17711-1-AP, 1:2000 for IF), GFP-tag (Proteintech, 50430-2-AP, 1:1000 for WB) and Myc-Tag (CST, 71D10, 1:1000 used for WB); goat polyclonal antibody against polycystin 2 (E-20, Santa Cruz, sc-10377, 1:500 for WB).

Techniques: Western Blot, Knockdown, Control, Staining

Journal: Cells

Article Title: The Immune Checkpoint Protein PD-L1 Regulates Ciliogenesis and Hedgehog Signaling

doi: 10.3390/cells13121003

Figure Lengend Snippet: PD-L1 interacts with BBS5 and PC-2. ( A ) Western blot analysis of the expression of PC-2, Ift140, BBS5, Rab8a and Ift20 in PD-L1 siRNA knockdown NIH3T3 cells compared to control siRNA cells. ( B ) Western blot analysis of the expression of PC-2, Ift140, BBS5, Rab8a and Ift20 in GFP-PD-L1 overexpressed NIH3T3 cells compared to GFP-vector control cells. ( C ) Western blot analysis of the expression of PC-2, IFT140, BBS5, RAB8a and IFT20 in PD-L1 siRNA knockdown RCTE cells compared to control siRNA cells. ( D ) Western blot of the co-immunoprecipitation analysis between GFP-PD-L1 and BBS5, and PC-2, in HEK293T cells. ( E ) Western blot of the co-immunoprecipitation analysis between Myc-PC-2 and BBS5 and Myc-PC-2 and GFP-PD-L1 in HEK293T cells. ( F ) Western blot analysis of the expression of PD-L1 and BBS5 in Myc-PC-2 overexpressed HEK293T cells.

Article Snippet: Primary antibodies used in this study are listed as follows: mouse monoclonal antibodies against acetylated α-tubulin (6-11B-1, Sigma, St. Louis, MO, USA, T7451, 1:4000 used for immunofluorescence [IF]), γ-tubulin (GTU-88, Sigma, T5326, 1:1000 used for IF), α-tubulin (DM1A, sc-32293, 1:1000), Ninein (F-7, Santa Cruz, Dallas, TX, USA, sc-390540, 1:300 for IF), Cep164 (E-9, Santa Cruz, sc-515403, 1:300 used for IF), C-nap1 (E-9, Santa Cruz, sc-515403, 1:300 used for IF), Smo (E-5, Santa Cruz, sc-166685, 1:100 used for IF), Gli1 (C-1, Santa Cruz, sc-515751, 1:500 used for WB), Giantin (9B6, Abcam, Waltham, MA, USA, ab37266, 1:500 used for IF), actin (AC-15, Sigma, A1978, 1:3000 used for Western blot [WB]), GFP-tag (B-2, sc-9996, 1:1000 for WB), and Myc (9E10, sc-40, 1:1000 for WB); rabbit polyclonal antibodies against PD-L1 (Proteintech, Rosemont, IL, USA, 17952-1-AP, 1:300 used for IF, and 1:1000 used for WB), Cep135 (Abcam, ab75005, 1:300 used for IF), Ift140 (Proteintech, 17460-1-AP, 1:300 used for IF, and 1:1000 for WB), Ift20 (Proteintech, 13615-1-AP, 1:300 used for IF, and 1:1000 for WB), Rab8a (Proteintech, 55296-1-AP, 1:300 used for IF, and 1:1000 for WB), BBS5 (Proteintech, 14569-1-AP, 1:300 used for IF, and 1:1000 for WB), ARL13B (Proteintech, 17711-1-AP, 1:2000 for IF), GFP-tag (Proteintech, 50430-2-AP, 1:1000 for WB) and Myc-Tag (CST, 71D10, 1:1000 used for WB); goat polyclonal antibody against polycystin 2 (E-20, Santa Cruz, sc-10377, 1:500 for WB).

Techniques: Western Blot, Expressing, Knockdown, Control, Plasmid Preparation, Immunoprecipitation

Journal: Cells

Article Title: The Immune Checkpoint Protein PD-L1 Regulates Ciliogenesis and Hedgehog Signaling

doi: 10.3390/cells13121003

Figure Lengend Snippet: PD-L1 regulates Hedgehog signal transduction. ( A ) Representative images of acetylated-α-tubulin (red) co-stained with Gli3 (green), in PD-L1 siRNA knockdown NIH3T3 cells without SAG stimulation, compared to control siRNA cells ( n > 100). All cells were counterstained with DAPI (blue). Scale bars, 20 μm. ( B ) Quantitative data of Gli3-positive cilia in unstimulated PD-L1 siRNA knockdown NIH3T3 cells compared to control siRNA cells. ( C ) Western blot analysis of the protein levels of Gli3 and Gli1 in unstimulated PD-L1 siRNA knockdown NIH3T3 cells compared to control siRNA cells. ( D ) qRT-PCR analysis of Hh signaling mediators (Gli1, Gli2 and Gli3) in unstimulated PD-L1 siRNA knockdown NIH3T3 cells compared to control siRNA cells. “ns” implies not significant.

Article Snippet: Primary antibodies used in this study are listed as follows: mouse monoclonal antibodies against acetylated α-tubulin (6-11B-1, Sigma, St. Louis, MO, USA, T7451, 1:4000 used for immunofluorescence [IF]), γ-tubulin (GTU-88, Sigma, T5326, 1:1000 used for IF), α-tubulin (DM1A, sc-32293, 1:1000), Ninein (F-7, Santa Cruz, Dallas, TX, USA, sc-390540, 1:300 for IF), Cep164 (E-9, Santa Cruz, sc-515403, 1:300 used for IF), C-nap1 (E-9, Santa Cruz, sc-515403, 1:300 used for IF), Smo (E-5, Santa Cruz, sc-166685, 1:100 used for IF), Gli1 (C-1, Santa Cruz, sc-515751, 1:500 used for WB), Giantin (9B6, Abcam, Waltham, MA, USA, ab37266, 1:500 used for IF), actin (AC-15, Sigma, A1978, 1:3000 used for Western blot [WB]), GFP-tag (B-2, sc-9996, 1:1000 for WB), and Myc (9E10, sc-40, 1:1000 for WB); rabbit polyclonal antibodies against PD-L1 (Proteintech, Rosemont, IL, USA, 17952-1-AP, 1:300 used for IF, and 1:1000 used for WB), Cep135 (Abcam, ab75005, 1:300 used for IF), Ift140 (Proteintech, 17460-1-AP, 1:300 used for IF, and 1:1000 for WB), Ift20 (Proteintech, 13615-1-AP, 1:300 used for IF, and 1:1000 for WB), Rab8a (Proteintech, 55296-1-AP, 1:300 used for IF, and 1:1000 for WB), BBS5 (Proteintech, 14569-1-AP, 1:300 used for IF, and 1:1000 for WB), ARL13B (Proteintech, 17711-1-AP, 1:2000 for IF), GFP-tag (Proteintech, 50430-2-AP, 1:1000 for WB) and Myc-Tag (CST, 71D10, 1:1000 used for WB); goat polyclonal antibody against polycystin 2 (E-20, Santa Cruz, sc-10377, 1:500 for WB).

Techniques: Transduction, Staining, Knockdown, Control, Western Blot, Quantitative RT-PCR

Journal: Cells

Article Title: The Immune Checkpoint Protein PD-L1 Regulates Ciliogenesis and Hedgehog Signaling

doi: 10.3390/cells13121003

Figure Lengend Snippet: Working model for PD-L1 in the regulation of ciliogenesis. Enriched expression of receptors on the ciliary membrane makes the primary cilium a specialized organelle for receiving and transducing extracellular stimuli into cells. Vesicles carrying ciliary proteins leave the Golgi and move toward the basal body of the primary cilium. Active forms of Rab8, a master modulator for the ciliary protein trafficking, and the BBSome complex, regulates the entry of protein cargo to the cilium. The activities and basal body localization of Rab8 are modulated by Golgi-associated Ift20 and the BBSome. Knockdown of PD-L1 enhances ciliogenesis by modulating the localization of proteins important for ciliary protein sorting, trafficking, and cilia sensory signaling. Depletion of PD-L1 increases cilia length and increases the ciliary recruitment of Rab8a and BBS5. Also, knockdown of PD-L1 increases the ciliary localization of cilia sensory receptor PC-2, and Gli3 which results in the repression of the Hh signaling pathway.

Article Snippet: Primary antibodies used in this study are listed as follows: mouse monoclonal antibodies against acetylated α-tubulin (6-11B-1, Sigma, St. Louis, MO, USA, T7451, 1:4000 used for immunofluorescence [IF]), γ-tubulin (GTU-88, Sigma, T5326, 1:1000 used for IF), α-tubulin (DM1A, sc-32293, 1:1000), Ninein (F-7, Santa Cruz, Dallas, TX, USA, sc-390540, 1:300 for IF), Cep164 (E-9, Santa Cruz, sc-515403, 1:300 used for IF), C-nap1 (E-9, Santa Cruz, sc-515403, 1:300 used for IF), Smo (E-5, Santa Cruz, sc-166685, 1:100 used for IF), Gli1 (C-1, Santa Cruz, sc-515751, 1:500 used for WB), Giantin (9B6, Abcam, Waltham, MA, USA, ab37266, 1:500 used for IF), actin (AC-15, Sigma, A1978, 1:3000 used for Western blot [WB]), GFP-tag (B-2, sc-9996, 1:1000 for WB), and Myc (9E10, sc-40, 1:1000 for WB); rabbit polyclonal antibodies against PD-L1 (Proteintech, Rosemont, IL, USA, 17952-1-AP, 1:300 used for IF, and 1:1000 used for WB), Cep135 (Abcam, ab75005, 1:300 used for IF), Ift140 (Proteintech, 17460-1-AP, 1:300 used for IF, and 1:1000 for WB), Ift20 (Proteintech, 13615-1-AP, 1:300 used for IF, and 1:1000 for WB), Rab8a (Proteintech, 55296-1-AP, 1:300 used for IF, and 1:1000 for WB), BBS5 (Proteintech, 14569-1-AP, 1:300 used for IF, and 1:1000 for WB), ARL13B (Proteintech, 17711-1-AP, 1:2000 for IF), GFP-tag (Proteintech, 50430-2-AP, 1:1000 for WB) and Myc-Tag (CST, 71D10, 1:1000 used for WB); goat polyclonal antibody against polycystin 2 (E-20, Santa Cruz, sc-10377, 1:500 for WB).

Techniques: Expressing, Membrane, Knockdown

Journal: Cell Communication and Signaling : CCS

Article Title: High expression of AMAP1, an ARF6 effector, is associated with elevated levels of PD-L1 and fibrosis of pancreatic cancer

doi: 10.1186/s12964-020-00608-8

Figure Lengend Snippet: High AMAP1 expression levels statistically correlate with PD-L1 expression in human PDACs and KPC tumors. a Representative IHC images of AMAP1 with IHC scores 0 to 3 and positive and negative staining of PD-L1 in clinical specimens, and their comparison with AMAP1 IHC scores. b IHC images and quantification of the PD-L1 staining of tumors formed by control (Irr) or AMAP1 -silenced ( shAMAP1 #1 and #2) KPC cells in C57BL/6 mice. Error bars represent the mean ± s.e.m. ** P < 0.01. c PD-L1 cell surface expression in IFNγ-treated or non-treated KPC cells, pretreated with shRNAs. MFI, median fluorescence intensity. Error bars represent the mean ± s.e.m. * P < 0.05, ** P < 0.01, *** P < 0.001. d Kaplan-Meier plots of the overall survival of patients with regard to PD-L1 positivity. P -values were obtained by t -tests ( a , b and c ) and by the log-rank test ( d ). Bars = 100 μm ( a and b )

Article Snippet: Other antibodies were purchased from commercial sources, as follows: rabbit monoclonal antibodies against PD-L1 (Cell Signaling) and phospho-FAK (Thermo Fisher Scientific), rabbit polyclonal antibody against PD-L1 (Novus), rabbit polyclonal antibody against collagen I (Proteintech), rabbit polyclonal antibody against FAK [ ], mouse monoclonal antibody against β-actin (Sigma-Aldrich).

Techniques: Expressing, Negative Staining, Comparison, Staining, Control, Fluorescence

Journal: Oncology Letters

Article Title: Prognostic impact of programmed cell death ligand 1 expression on long-term oncologic outcomes in colorectal cancer

doi: 10.3892/ol.2018.9264

Figure Lengend Snippet: (A) High (52.8%) and (B) low (47.2%) PD-L1 expression.

Article Snippet: The primary antibody was a rabbit polyclonal antibody against PD-L1 (1:400, Anti-PD-L1; AnaSpec, Fremont, CA, USA).

Techniques: Expressing

Journal: Oncology Letters

Article Title: Prognostic impact of programmed cell death ligand 1 expression on long-term oncologic outcomes in colorectal cancer

doi: 10.3892/ol.2018.9264

Figure Lengend Snippet: Ten-year overall and disease-free survival of patients with CRC according to PD-L1 expression. The 10-year OS and DFS rates were significantly higher in the high PD-L1 expression group than in the low PD-L1 expression group and low PD-L1 expression was significantly associated with tumor relapse and poor prognosis in stage III CRC. CRC, colorectal cancer.

Article Snippet: The primary antibody was a rabbit polyclonal antibody against PD-L1 (1:400, Anti-PD-L1; AnaSpec, Fremont, CA, USA).

Techniques: Expressing

Journal: Oncology Letters

Article Title: Prognostic impact of programmed cell death ligand 1 expression on long-term oncologic outcomes in colorectal cancer

doi: 10.3892/ol.2018.9264

Figure Lengend Snippet: Prognostic factors of survival in univariate analysis.

Article Snippet: The primary antibody was a rabbit polyclonal antibody against PD-L1 (1:400, Anti-PD-L1; AnaSpec, Fremont, CA, USA).

Techniques: Expressing

Journal: Oncology Letters

Article Title: Prognostic impact of programmed cell death ligand 1 expression on long-term oncologic outcomes in colorectal cancer

doi: 10.3892/ol.2018.9264

Figure Lengend Snippet: Ten-year overall and disease-free survival of patients with CRC according to prognostic variables with statistical significance. Univariate analyses revealed that T-stage, N-stage, tumor differentiation, lymphovascular invasion, PD-L1 expression, and perinodal extension were significantly associated with overall and disease-free survival. CRC, colorectal cancer.

Article Snippet: The primary antibody was a rabbit polyclonal antibody against PD-L1 (1:400, Anti-PD-L1; AnaSpec, Fremont, CA, USA).

Techniques: Expressing