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human rat cd44 5640s  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc human rat cd44 5640s
    Identification and distribution <t>of</t> <t>CD44-positive</t> cancer stem-like cells in human pNET tissues. A . Human pNET specimens in patient 1 were co-stained with <t>CD44</t> and α-SMA antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (blue) by immunofluorescence microscopy. CD44-positive cancer cells (red) marked by yellow arrowheads were close to the α-SMA positive (green) blood vessels. B : Human pNET specimens in patient 2 were co-stained as in A . CD44-positive cancer cells (green) were indicated by yellow arrowheads, which were close to the α-SMA positive (red) blood vessels. Note: A group of CD44-positive cells appear to distribute within the α-SMA-positive vascular niche or nearby blood vessels. Shown are representative images from different individual patients. Bar = 10 or 20 µm. C . Human pNET tissues were co-stained with CD44 antibodies and CD45 antibodies followed by appropriate secondary antibodies. Overlay images were collected by immunofluorescence microscopy. CD44-positive (green, yellow arrowheads) and both CD44-positive and CD45-positive cells (orange, blue arrowheads) were observed under a fluorescence microscope. Images were acquired by a fluorescence microscope equipped with a CCD camera. Shown are representative images from two individual patients. Bar = 10 µm. Note: Cells positive with both CD44 and CD45 are smaller and regarded as lymphatic cells (orange), a subset of cancer cells are bigger and positive for CD44 but negative for CD45, indicating their stem-like phenotype.
    Human Rat Cd44 5640s, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 212 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/human+rat+cd44+5640s/CD44+Mouse+mAb/bio_rxiv__2022__02__17__480869-37-18-26
    Average 95 stars, based on 212 article reviews
    human rat cd44 5640s - by Bioz Stars, 2026-10
    95/100 stars

    Images

    1) Product Images from "PKD-1 Signaling Is Required for the Maintenance of CSCs with Epithelial-mesenchymal Plasticity in Pancreatic Neuroendocrine Tumors"

    Article Title: PKD-1 Signaling Is Required for the Maintenance of CSCs with Epithelial-mesenchymal Plasticity in Pancreatic Neuroendocrine Tumors

    Journal: bioRxiv

    doi: 10.1101/2022.02.17.480869

    Identification and distribution of CD44-positive cancer stem-like cells in human pNET tissues. A . Human pNET specimens in patient 1 were co-stained with CD44 and α-SMA antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (blue) by immunofluorescence microscopy. CD44-positive cancer cells (red) marked by yellow arrowheads were close to the α-SMA positive (green) blood vessels. B : Human pNET specimens in patient 2 were co-stained as in A . CD44-positive cancer cells (green) were indicated by yellow arrowheads, which were close to the α-SMA positive (red) blood vessels. Note: A group of CD44-positive cells appear to distribute within the α-SMA-positive vascular niche or nearby blood vessels. Shown are representative images from different individual patients. Bar = 10 or 20 µm. C . Human pNET tissues were co-stained with CD44 antibodies and CD45 antibodies followed by appropriate secondary antibodies. Overlay images were collected by immunofluorescence microscopy. CD44-positive (green, yellow arrowheads) and both CD44-positive and CD45-positive cells (orange, blue arrowheads) were observed under a fluorescence microscope. Images were acquired by a fluorescence microscope equipped with a CCD camera. Shown are representative images from two individual patients. Bar = 10 µm. Note: Cells positive with both CD44 and CD45 are smaller and regarded as lymphatic cells (orange), a subset of cancer cells are bigger and positive for CD44 but negative for CD45, indicating their stem-like phenotype.
    Figure Legend Snippet: Identification and distribution of CD44-positive cancer stem-like cells in human pNET tissues. A . Human pNET specimens in patient 1 were co-stained with CD44 and α-SMA antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (blue) by immunofluorescence microscopy. CD44-positive cancer cells (red) marked by yellow arrowheads were close to the α-SMA positive (green) blood vessels. B : Human pNET specimens in patient 2 were co-stained as in A . CD44-positive cancer cells (green) were indicated by yellow arrowheads, which were close to the α-SMA positive (red) blood vessels. Note: A group of CD44-positive cells appear to distribute within the α-SMA-positive vascular niche or nearby blood vessels. Shown are representative images from different individual patients. Bar = 10 or 20 µm. C . Human pNET tissues were co-stained with CD44 antibodies and CD45 antibodies followed by appropriate secondary antibodies. Overlay images were collected by immunofluorescence microscopy. CD44-positive (green, yellow arrowheads) and both CD44-positive and CD45-positive cells (orange, blue arrowheads) were observed under a fluorescence microscope. Images were acquired by a fluorescence microscope equipped with a CCD camera. Shown are representative images from two individual patients. Bar = 10 µm. Note: Cells positive with both CD44 and CD45 are smaller and regarded as lymphatic cells (orange), a subset of cancer cells are bigger and positive for CD44 but negative for CD45, indicating their stem-like phenotype.

    Techniques Used: Staining, Immunofluorescence, Microscopy, Fluorescence

    Stem-like phenotype of cancer cells in pNETs. A . A human pancreatic tissue control (top panel) and human pNET specimens from two individual patients (middle and lower panels) were co-stained with CD44 and α-SMA antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (blue) by immunofluorescence microscopy. CD44-positive cancer stem-like cells were marked by yellow arrowheads, which were close to the α-SMA positive (green, red arrowheads) blood vessels. The fluorescence images were acquired by an immunofluorescence microscope equipped with a CCD camera, and representative images are shown, along with H & E staining for tumor tissue structures. Bar = 10 µm.
    Figure Legend Snippet: Stem-like phenotype of cancer cells in pNETs. A . A human pancreatic tissue control (top panel) and human pNET specimens from two individual patients (middle and lower panels) were co-stained with CD44 and α-SMA antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (blue) by immunofluorescence microscopy. CD44-positive cancer stem-like cells were marked by yellow arrowheads, which were close to the α-SMA positive (green, red arrowheads) blood vessels. The fluorescence images were acquired by an immunofluorescence microscope equipped with a CCD camera, and representative images are shown, along with H & E staining for tumor tissue structures. Bar = 10 µm.

    Techniques Used: Control, Staining, Immunofluorescence, Microscopy, Fluorescence

    PKD-1 signaling in the maintenance of cancer stem-like features in pNETs. A . Distribution of PKD-1 + and CD44 + CSCs within the vascular niche. Human pNET specimens were co-stained with CD44 and PKD-1 antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (Blue). Stem-like cells with CD44-positive (green), PKD-1-positive (red) or both positive (pink) were observed under a fluorescence microscope. A few CD44-positive cancer stem-like cells tended to accumulate near the vascular lumen (red arrow heads), and PKD-1-positive CSCs might be leaving tumor nests (stars) for the vascular lumen. The fluorescence images were acquired by an immunofluorescence microscope equipped with a CCD camera. Shown are representative images. Bar = 10 µm. B . BON and QGP-1 cells were transfected with siRNA control and siPKD-1 to knock down PKD-1. Knockdown efficiency was confirmed by Western Blots (upper panel). The control and BON cells with knocked-down PKD-1 were subjected to tumorsphere formation assays. Images were acquired by the OLYMPUS CK30 microscope. Representative images are shown for tumorsphere formation (lower panel). C . Cell lysates were extracted from BON and QGP-1 cells exposed to the vehicle control, 10 μ M LPA, 2 μ M CRT0066101, or their combinations after 24 hours. The expression levels of phosphorylated PKD-1 and PKD-1 were detected by Western blots. Shown are representative images. D . BON and QGP-1 cells were exposed to 10 μ M LPA, 2 μ M CRT0066101, or their combination for 24 hours, and total RNA was extracted for the detection of mRNA levels of genes related to stemness properties by RT-qPCR. E . Effect of PKD inhibitor in tumorsphere formation. BON cells were cultured in complete MammoCult(tm) medium with the treatment of 10 μ M LPA, 2 μ M CRT0066101, or their combination for 7 days. The mammary spheres were counted under the OLYMPUS CK30 microscope. F . Control and BON cells with PKD-1 knockdown were exposed to 10 μ M LPA, 2 μ M CRT0066101, or their combination for 24 hours, and total RNA was extracted for the detection of mRNA levels of genes related to stemness properties by RT-qPCR. Note: Triplicate experiments were performed, and the results are shown as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.
    Figure Legend Snippet: PKD-1 signaling in the maintenance of cancer stem-like features in pNETs. A . Distribution of PKD-1 + and CD44 + CSCs within the vascular niche. Human pNET specimens were co-stained with CD44 and PKD-1 antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (Blue). Stem-like cells with CD44-positive (green), PKD-1-positive (red) or both positive (pink) were observed under a fluorescence microscope. A few CD44-positive cancer stem-like cells tended to accumulate near the vascular lumen (red arrow heads), and PKD-1-positive CSCs might be leaving tumor nests (stars) for the vascular lumen. The fluorescence images were acquired by an immunofluorescence microscope equipped with a CCD camera. Shown are representative images. Bar = 10 µm. B . BON and QGP-1 cells were transfected with siRNA control and siPKD-1 to knock down PKD-1. Knockdown efficiency was confirmed by Western Blots (upper panel). The control and BON cells with knocked-down PKD-1 were subjected to tumorsphere formation assays. Images were acquired by the OLYMPUS CK30 microscope. Representative images are shown for tumorsphere formation (lower panel). C . Cell lysates were extracted from BON and QGP-1 cells exposed to the vehicle control, 10 μ M LPA, 2 μ M CRT0066101, or their combinations after 24 hours. The expression levels of phosphorylated PKD-1 and PKD-1 were detected by Western blots. Shown are representative images. D . BON and QGP-1 cells were exposed to 10 μ M LPA, 2 μ M CRT0066101, or their combination for 24 hours, and total RNA was extracted for the detection of mRNA levels of genes related to stemness properties by RT-qPCR. E . Effect of PKD inhibitor in tumorsphere formation. BON cells were cultured in complete MammoCult(tm) medium with the treatment of 10 μ M LPA, 2 μ M CRT0066101, or their combination for 7 days. The mammary spheres were counted under the OLYMPUS CK30 microscope. F . Control and BON cells with PKD-1 knockdown were exposed to 10 μ M LPA, 2 μ M CRT0066101, or their combination for 24 hours, and total RNA was extracted for the detection of mRNA levels of genes related to stemness properties by RT-qPCR. Note: Triplicate experiments were performed, and the results are shown as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

    Techniques Used: Staining, Fluorescence, Microscopy, Immunofluorescence, Transfection, Control, Knockdown, Western Blot, Expressing, Quantitative RT-PCR, Cell Culture

    Distribution of CD44-and/or PKD-1-positive cancer stem-like cells in pNET tissues. Human pNET specimens were co-stained with CD44 antibodies and PKD-1 antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (blue). CD44-positive (green), PKD-1-positive (red) or both positive were observed under a fluorescence microscope. Cancer cells with high levels of both CD44 and PKD-1 (yellow arrowheads) likely left the tumor nests (white stars) and accumulated in the nearby vascular lumen (red arrowheads). Fluorescence images were acquired by an immunofluorescence microscope equipped with a CCD camera. Shown are representative images from two individual patients. Bar = 10 µm. Note: These are additional pictures for .
    Figure Legend Snippet: Distribution of CD44-and/or PKD-1-positive cancer stem-like cells in pNET tissues. Human pNET specimens were co-stained with CD44 antibodies and PKD-1 antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (blue). CD44-positive (green), PKD-1-positive (red) or both positive were observed under a fluorescence microscope. Cancer cells with high levels of both CD44 and PKD-1 (yellow arrowheads) likely left the tumor nests (white stars) and accumulated in the nearby vascular lumen (red arrowheads). Fluorescence images were acquired by an immunofluorescence microscope equipped with a CCD camera. Shown are representative images from two individual patients. Bar = 10 µm. Note: These are additional pictures for .

    Techniques Used: Staining, Fluorescence, Microscopy, Immunofluorescence

    Related Articles

    Immunostaining:

    Article Title: LPA/PKD-1 signaling promotes development of arteriolar niche that supports self-renewal of breast cancer stem cells and stemness
    Article Snippet: Antibodies include PKD/PKCμ (D4J1N), Neurophilin1 (D62C7), phospho-Erk (1/2) (D13.14.4E), and human/mouse Erk (1/2) (137F5) (Cell Signaling Technology), ALDH1A1 (AF5869) (R&D Systems) and EphrinB2 (ab131536) and CD36 (ab133625) (Abcam). .. Antibodies for immunostaining include rabbit anti-human/mouse PKD-1 (4502371) and mouse anti-mouse α-smooth muscle actin(A2547) (Sigma-Aldrich), mouse anti-human/rat CD44 (5640S) and rabbit anti-human/mouse ALDH1A1 (12035S) (Cell Signaling Technology), rabbit anti-human/mouse CD36 (ab133625) (Abcam), AlexaFluor 594 conjugated donkey anti-rabbit IgG (A21207), AlexaFluor 594 conjugated donkey anti-mouse IgG(A21203), AlexaFluor 594 conjugated donkey anti-goat IgG(A11058), AlexaFluor 488 conjugated donkey anti-goat IgG(A11055), AlexaFluor 488 conjugated donkey anti-rabbit IgG(A21206), and AlexaFluor 488 conjugated goat anti-mouse IgG(A11001) (Invitrogen). ..

    Article Title: Development of arteriolar niche and self-renewal of breast cancer stem cells by lysophosphatidic Acid/protein kinase D signaling&nbsp;
    Article Snippet: Antibodies include PKD/PKCμ (D4J1N), Neurophilin1 (D62C7), phospho-Erk (1/2) (D13.14.4E), and human/mouse Erk (1/2) (137F5) (Cell Signaling Technology), ALDH1A1 (AF5869) (R&D Systems) and ephrinB2 (ab131536) and CD36 (ab133625) (Abcam). .. Antibodies for immunostaining include rabbit anti-human/mouse PKD-1 (4502371) and mouse anti-mouse α-smooth muscle actin(A2547) (SigmaAldrich), mouse anti-human/rat CD44 (5640S) and rabbit anti-human/mouse ALDH1A1 (12035S) (Cell Signaling Technology), rabbit anti-human/mouse CD36 (ab133625) (Abcam), AlexaFluor 594 conjugated donkey anti-rabbit IgG (A21207), AlexaFluor 594 conjugated donkey anti-mouse IgG(A21203), AlexaFluor 594 conjugated donkey anti-goat IgG(A11058), AlexaFluor 488 conjugated donkey anti-goat IgG(A11055), AlexaFluor 488 conjugated donkey anti-rabbit IgG(A21206), and AlexaFluor 488 conjugated goat antimouse IgG(A11001) (Invitrogen). ..

    Article Title: Development of an arteriolar niche and self-renewal of breast cancer stem cells by lysophosphatidic acid/protein kinase D signaling
    Article Snippet: Antibodies include PKD/PKCμ (D4J1N), Neurophilin1 (D62C7), phospho-Erk (1/2) (D13.14.4E), and human/mouse Erk (1/2) (137F5) (Cell Signaling Technology), ALDH1A1 (AF5869) (R&D Systems) and ephrinB2 (ab131536) and CD36 (ab133625) (Abcam). .. Antibodies for immunostaining include rabbit anti-human/mouse PKD-1 (4502371) and mouse anti-mouse α-smooth muscle actin(A2547) (Sigma-Aldrich), mouse anti-human/rat CD44 (5640S) and rabbit anti-human/mouse ALDH1A1 (12035S) (Cell Signaling Technology), rabbit anti-human/mouse CD36 (ab133625) (Abcam), AlexaFluor 594 conjugated donkey anti-rabbit IgG (A21207), AlexaFluor 594 conjugated donkey anti-mouse IgG(A21203), AlexaFluor 594 conjugated donkey anti-goat IgG(A11058), AlexaFluor 488 conjugated donkey anti-goat IgG(A11055), AlexaFluor 488 conjugated donkey anti-rabbit IgG(A21206), AlexaFluor 350 conjugated goat anti-mouse IgG (H + L, A11045), and AlexaFluor 488 conjugated goat anti-mouse IgG(A11001) (Invitrogen). ..

    Article Title: PKD-1 Signaling Is Required for the Maintenance of CSCs with Epithelial-mesenchymal Plasticity in Pancreatic Neuroendocrine Tumors
    Article Snippet: Antibodies for immunoblot included PKD/PKCμ (90039S), phospho-PKD/PKCμ (2051T), phospho-Erk (1/2) (4370), Erk (1/2) (137F5), E-cadherin (24E10), vimentin (D21H3) (Cell Signaling Technology) and ALDH1A1 (AF5869) (R&D Systems). .. Antibodies for immunostaining included rabbit anti-human/mouse PKD-1 (SAB4502371) and mouse anti-human/mouse α-smooth muscle actin (α-SMA, A2547) (Sigma-Aldrich), mouse anti-human/rat CD44 (5640S) and HRP-linked anti-rabbit IgG (7074) (Cell Signaling Technology), CD45 monoclonal Antibody (14-9457-80), AlexaFluor 594 conjugated donkey anti-rabbit IgG (A21207), AlexaFluor 594 conjugated donkey anti-mouse IgG(A21203), AlexaFluor 594 conjugated donkey anti-goat IgG (A11058), AlexaFluor 488 conjugated donkey anti-goat IgG(A11055), AlexaFluor 488 conjugated donkey anti-rabbit IgG (A21206), and AlexaFluor 488 conjugated goat anti-mouse IgG (A11001) (Invitrogen). ..



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    human rat cd44 5640s  (Cell Signaling Technology Inc)
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    Cell Signaling Technology Inc human rat cd44 5640s
    Identification and distribution <t>of</t> <t>CD44-positive</t> cancer stem-like cells in human pNET tissues. A . Human pNET specimens in patient 1 were co-stained with <t>CD44</t> and α-SMA antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (blue) by immunofluorescence microscopy. CD44-positive cancer cells (red) marked by yellow arrowheads were close to the α-SMA positive (green) blood vessels. B : Human pNET specimens in patient 2 were co-stained as in A . CD44-positive cancer cells (green) were indicated by yellow arrowheads, which were close to the α-SMA positive (red) blood vessels. Note: A group of CD44-positive cells appear to distribute within the α-SMA-positive vascular niche or nearby blood vessels. Shown are representative images from different individual patients. Bar = 10 or 20 µm. C . Human pNET tissues were co-stained with CD44 antibodies and CD45 antibodies followed by appropriate secondary antibodies. Overlay images were collected by immunofluorescence microscopy. CD44-positive (green, yellow arrowheads) and both CD44-positive and CD45-positive cells (orange, blue arrowheads) were observed under a fluorescence microscope. Images were acquired by a fluorescence microscope equipped with a CCD camera. Shown are representative images from two individual patients. Bar = 10 µm. Note: Cells positive with both CD44 and CD45 are smaller and regarded as lymphatic cells (orange), a subset of cancer cells are bigger and positive for CD44 but negative for CD45, indicating their stem-like phenotype.
    Human Rat Cd44 5640s, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/human+rat+cd44+5640s/CD44+Mouse+mAb/bio_rxiv__2022__02__17__480869-37-18-26
    Average 95 stars, based on 1 article reviews
    human rat cd44 5640s - by Bioz Stars, 2026-10
    95/100 stars
    		  Buy from Supplier

    Image Search Results


    Identification and distribution of CD44-positive cancer stem-like cells in human pNET tissues. A . Human pNET specimens in patient 1 were co-stained with CD44 and α-SMA antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (blue) by immunofluorescence microscopy. CD44-positive cancer cells (red) marked by yellow arrowheads were close to the α-SMA positive (green) blood vessels. B : Human pNET specimens in patient 2 were co-stained as in A . CD44-positive cancer cells (green) were indicated by yellow arrowheads, which were close to the α-SMA positive (red) blood vessels. Note: A group of CD44-positive cells appear to distribute within the α-SMA-positive vascular niche or nearby blood vessels. Shown are representative images from different individual patients. Bar = 10 or 20 µm. C . Human pNET tissues were co-stained with CD44 antibodies and CD45 antibodies followed by appropriate secondary antibodies. Overlay images were collected by immunofluorescence microscopy. CD44-positive (green, yellow arrowheads) and both CD44-positive and CD45-positive cells (orange, blue arrowheads) were observed under a fluorescence microscope. Images were acquired by a fluorescence microscope equipped with a CCD camera. Shown are representative images from two individual patients. Bar = 10 µm. Note: Cells positive with both CD44 and CD45 are smaller and regarded as lymphatic cells (orange), a subset of cancer cells are bigger and positive for CD44 but negative for CD45, indicating their stem-like phenotype.

    Journal: bioRxiv

    Article Title: PKD-1 Signaling Is Required for the Maintenance of CSCs with Epithelial-mesenchymal Plasticity in Pancreatic Neuroendocrine Tumors

    doi: 10.1101/2022.02.17.480869

    Figure Lengend Snippet: Identification and distribution of CD44-positive cancer stem-like cells in human pNET tissues. A . Human pNET specimens in patient 1 were co-stained with CD44 and α-SMA antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (blue) by immunofluorescence microscopy. CD44-positive cancer cells (red) marked by yellow arrowheads were close to the α-SMA positive (green) blood vessels. B : Human pNET specimens in patient 2 were co-stained as in A . CD44-positive cancer cells (green) were indicated by yellow arrowheads, which were close to the α-SMA positive (red) blood vessels. Note: A group of CD44-positive cells appear to distribute within the α-SMA-positive vascular niche or nearby blood vessels. Shown are representative images from different individual patients. Bar = 10 or 20 µm. C . Human pNET tissues were co-stained with CD44 antibodies and CD45 antibodies followed by appropriate secondary antibodies. Overlay images were collected by immunofluorescence microscopy. CD44-positive (green, yellow arrowheads) and both CD44-positive and CD45-positive cells (orange, blue arrowheads) were observed under a fluorescence microscope. Images were acquired by a fluorescence microscope equipped with a CCD camera. Shown are representative images from two individual patients. Bar = 10 µm. Note: Cells positive with both CD44 and CD45 are smaller and regarded as lymphatic cells (orange), a subset of cancer cells are bigger and positive for CD44 but negative for CD45, indicating their stem-like phenotype.

    Article Snippet: Antibodies for immunostaining included rabbit anti-human/mouse PKD-1 (SAB4502371) and mouse anti-human/mouse α-smooth muscle actin (α-SMA, A2547) (Sigma-Aldrich), mouse anti-human/rat CD44 (5640S) and HRP-linked anti-rabbit IgG (7074) (Cell Signaling Technology), CD45 monoclonal Antibody (14-9457-80), AlexaFluor 594 conjugated donkey anti-rabbit IgG (A21207), AlexaFluor 594 conjugated donkey anti-mouse IgG(A21203), AlexaFluor 594 conjugated donkey anti-goat IgG (A11058), AlexaFluor 488 conjugated donkey anti-goat IgG(A11055), AlexaFluor 488 conjugated donkey anti-rabbit IgG (A21206), and AlexaFluor 488 conjugated goat anti-mouse IgG (A11001) (Invitrogen).

    Techniques: Staining, Immunofluorescence, Microscopy, Fluorescence

    Stem-like phenotype of cancer cells in pNETs. A . A human pancreatic tissue control (top panel) and human pNET specimens from two individual patients (middle and lower panels) were co-stained with CD44 and α-SMA antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (blue) by immunofluorescence microscopy. CD44-positive cancer stem-like cells were marked by yellow arrowheads, which were close to the α-SMA positive (green, red arrowheads) blood vessels. The fluorescence images were acquired by an immunofluorescence microscope equipped with a CCD camera, and representative images are shown, along with H & E staining for tumor tissue structures. Bar = 10 µm.

    Journal: bioRxiv

    Article Title: PKD-1 Signaling Is Required for the Maintenance of CSCs with Epithelial-mesenchymal Plasticity in Pancreatic Neuroendocrine Tumors

    doi: 10.1101/2022.02.17.480869

    Figure Lengend Snippet: Stem-like phenotype of cancer cells in pNETs. A . A human pancreatic tissue control (top panel) and human pNET specimens from two individual patients (middle and lower panels) were co-stained with CD44 and α-SMA antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (blue) by immunofluorescence microscopy. CD44-positive cancer stem-like cells were marked by yellow arrowheads, which were close to the α-SMA positive (green, red arrowheads) blood vessels. The fluorescence images were acquired by an immunofluorescence microscope equipped with a CCD camera, and representative images are shown, along with H & E staining for tumor tissue structures. Bar = 10 µm.

    Article Snippet: Antibodies for immunostaining included rabbit anti-human/mouse PKD-1 (SAB4502371) and mouse anti-human/mouse α-smooth muscle actin (α-SMA, A2547) (Sigma-Aldrich), mouse anti-human/rat CD44 (5640S) and HRP-linked anti-rabbit IgG (7074) (Cell Signaling Technology), CD45 monoclonal Antibody (14-9457-80), AlexaFluor 594 conjugated donkey anti-rabbit IgG (A21207), AlexaFluor 594 conjugated donkey anti-mouse IgG(A21203), AlexaFluor 594 conjugated donkey anti-goat IgG (A11058), AlexaFluor 488 conjugated donkey anti-goat IgG(A11055), AlexaFluor 488 conjugated donkey anti-rabbit IgG (A21206), and AlexaFluor 488 conjugated goat anti-mouse IgG (A11001) (Invitrogen).

    Techniques: Control, Staining, Immunofluorescence, Microscopy, Fluorescence

    PKD-1 signaling in the maintenance of cancer stem-like features in pNETs. A . Distribution of PKD-1 + and CD44 + CSCs within the vascular niche. Human pNET specimens were co-stained with CD44 and PKD-1 antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (Blue). Stem-like cells with CD44-positive (green), PKD-1-positive (red) or both positive (pink) were observed under a fluorescence microscope. A few CD44-positive cancer stem-like cells tended to accumulate near the vascular lumen (red arrow heads), and PKD-1-positive CSCs might be leaving tumor nests (stars) for the vascular lumen. The fluorescence images were acquired by an immunofluorescence microscope equipped with a CCD camera. Shown are representative images. Bar = 10 µm. B . BON and QGP-1 cells were transfected with siRNA control and siPKD-1 to knock down PKD-1. Knockdown efficiency was confirmed by Western Blots (upper panel). The control and BON cells with knocked-down PKD-1 were subjected to tumorsphere formation assays. Images were acquired by the OLYMPUS CK30 microscope. Representative images are shown for tumorsphere formation (lower panel). C . Cell lysates were extracted from BON and QGP-1 cells exposed to the vehicle control, 10 μ M LPA, 2 μ M CRT0066101, or their combinations after 24 hours. The expression levels of phosphorylated PKD-1 and PKD-1 were detected by Western blots. Shown are representative images. D . BON and QGP-1 cells were exposed to 10 μ M LPA, 2 μ M CRT0066101, or their combination for 24 hours, and total RNA was extracted for the detection of mRNA levels of genes related to stemness properties by RT-qPCR. E . Effect of PKD inhibitor in tumorsphere formation. BON cells were cultured in complete MammoCult(tm) medium with the treatment of 10 μ M LPA, 2 μ M CRT0066101, or their combination for 7 days. The mammary spheres were counted under the OLYMPUS CK30 microscope. F . Control and BON cells with PKD-1 knockdown were exposed to 10 μ M LPA, 2 μ M CRT0066101, or their combination for 24 hours, and total RNA was extracted for the detection of mRNA levels of genes related to stemness properties by RT-qPCR. Note: Triplicate experiments were performed, and the results are shown as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

    Journal: bioRxiv

    Article Title: PKD-1 Signaling Is Required for the Maintenance of CSCs with Epithelial-mesenchymal Plasticity in Pancreatic Neuroendocrine Tumors

    doi: 10.1101/2022.02.17.480869

    Figure Lengend Snippet: PKD-1 signaling in the maintenance of cancer stem-like features in pNETs. A . Distribution of PKD-1 + and CD44 + CSCs within the vascular niche. Human pNET specimens were co-stained with CD44 and PKD-1 antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (Blue). Stem-like cells with CD44-positive (green), PKD-1-positive (red) or both positive (pink) were observed under a fluorescence microscope. A few CD44-positive cancer stem-like cells tended to accumulate near the vascular lumen (red arrow heads), and PKD-1-positive CSCs might be leaving tumor nests (stars) for the vascular lumen. The fluorescence images were acquired by an immunofluorescence microscope equipped with a CCD camera. Shown are representative images. Bar = 10 µm. B . BON and QGP-1 cells were transfected with siRNA control and siPKD-1 to knock down PKD-1. Knockdown efficiency was confirmed by Western Blots (upper panel). The control and BON cells with knocked-down PKD-1 were subjected to tumorsphere formation assays. Images were acquired by the OLYMPUS CK30 microscope. Representative images are shown for tumorsphere formation (lower panel). C . Cell lysates were extracted from BON and QGP-1 cells exposed to the vehicle control, 10 μ M LPA, 2 μ M CRT0066101, or their combinations after 24 hours. The expression levels of phosphorylated PKD-1 and PKD-1 were detected by Western blots. Shown are representative images. D . BON and QGP-1 cells were exposed to 10 μ M LPA, 2 μ M CRT0066101, or their combination for 24 hours, and total RNA was extracted for the detection of mRNA levels of genes related to stemness properties by RT-qPCR. E . Effect of PKD inhibitor in tumorsphere formation. BON cells were cultured in complete MammoCult(tm) medium with the treatment of 10 μ M LPA, 2 μ M CRT0066101, or their combination for 7 days. The mammary spheres were counted under the OLYMPUS CK30 microscope. F . Control and BON cells with PKD-1 knockdown were exposed to 10 μ M LPA, 2 μ M CRT0066101, or their combination for 24 hours, and total RNA was extracted for the detection of mRNA levels of genes related to stemness properties by RT-qPCR. Note: Triplicate experiments were performed, and the results are shown as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

    Article Snippet: Antibodies for immunostaining included rabbit anti-human/mouse PKD-1 (SAB4502371) and mouse anti-human/mouse α-smooth muscle actin (α-SMA, A2547) (Sigma-Aldrich), mouse anti-human/rat CD44 (5640S) and HRP-linked anti-rabbit IgG (7074) (Cell Signaling Technology), CD45 monoclonal Antibody (14-9457-80), AlexaFluor 594 conjugated donkey anti-rabbit IgG (A21207), AlexaFluor 594 conjugated donkey anti-mouse IgG(A21203), AlexaFluor 594 conjugated donkey anti-goat IgG (A11058), AlexaFluor 488 conjugated donkey anti-goat IgG(A11055), AlexaFluor 488 conjugated donkey anti-rabbit IgG (A21206), and AlexaFluor 488 conjugated goat anti-mouse IgG (A11001) (Invitrogen).

    Techniques: Staining, Fluorescence, Microscopy, Immunofluorescence, Transfection, Control, Knockdown, Western Blot, Expressing, Quantitative RT-PCR, Cell Culture

    Distribution of CD44-and/or PKD-1-positive cancer stem-like cells in pNET tissues. Human pNET specimens were co-stained with CD44 antibodies and PKD-1 antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (blue). CD44-positive (green), PKD-1-positive (red) or both positive were observed under a fluorescence microscope. Cancer cells with high levels of both CD44 and PKD-1 (yellow arrowheads) likely left the tumor nests (white stars) and accumulated in the nearby vascular lumen (red arrowheads). Fluorescence images were acquired by an immunofluorescence microscope equipped with a CCD camera. Shown are representative images from two individual patients. Bar = 10 µm. Note: These are additional pictures for .

    Journal: bioRxiv

    Article Title: PKD-1 Signaling Is Required for the Maintenance of CSCs with Epithelial-mesenchymal Plasticity in Pancreatic Neuroendocrine Tumors

    doi: 10.1101/2022.02.17.480869

    Figure Lengend Snippet: Distribution of CD44-and/or PKD-1-positive cancer stem-like cells in pNET tissues. Human pNET specimens were co-stained with CD44 antibodies and PKD-1 antibodies followed by appropriate secondary antibodies, with DAPI staining the nuclei (blue). CD44-positive (green), PKD-1-positive (red) or both positive were observed under a fluorescence microscope. Cancer cells with high levels of both CD44 and PKD-1 (yellow arrowheads) likely left the tumor nests (white stars) and accumulated in the nearby vascular lumen (red arrowheads). Fluorescence images were acquired by an immunofluorescence microscope equipped with a CCD camera. Shown are representative images from two individual patients. Bar = 10 µm. Note: These are additional pictures for .

    Article Snippet: Antibodies for immunostaining included rabbit anti-human/mouse PKD-1 (SAB4502371) and mouse anti-human/mouse α-smooth muscle actin (α-SMA, A2547) (Sigma-Aldrich), mouse anti-human/rat CD44 (5640S) and HRP-linked anti-rabbit IgG (7074) (Cell Signaling Technology), CD45 monoclonal Antibody (14-9457-80), AlexaFluor 594 conjugated donkey anti-rabbit IgG (A21207), AlexaFluor 594 conjugated donkey anti-mouse IgG(A21203), AlexaFluor 594 conjugated donkey anti-goat IgG (A11058), AlexaFluor 488 conjugated donkey anti-goat IgG(A11055), AlexaFluor 488 conjugated donkey anti-rabbit IgG (A21206), and AlexaFluor 488 conjugated goat anti-mouse IgG (A11001) (Invitrogen).

    Techniques: Staining, Fluorescence, Microscopy, Immunofluorescence