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sw1353  (ATCC)


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    Structured Review

    ATCC sw1353
    Mitochondrial dysfunction activates ADAMTS-5 expression in chondrocytes (A–D) The impact of inhibiting mitochondrial respiratory chain with oligo A (30 μg/mL) on mitochondrial membrane potential (A, n = 6), ROS generation (B, n = 6), cell survival (C, n = 6), and the mRNA expression of IFNB1 and ISGs (D, n = 6) in the <t>SW1353</t> cell line. Scale bars in (B) are 20 μm. (E) Immunoblotting was used to analyze the phosphorylation of PKR and eIF2α following oligo A treatment in the SW1353 cell line, with quantification displayed on the right ( n = 6). (F and G) RT-qPCR and immunoblotting were conducted to assess the expression of ADAMTS-5 mRNA (F, n = 6) and protein (G) in response to oligo A treatment in the SW1353 cell line, with immunoblotting quantification shown on the right ( n = 6). Data (mean ± std) was representative of three independent experiments. An unpaired student’s t test was conducted to compare two groups. ∗, p < 0.05, ∗, p < 0.01, ∗∗∗, p < 0.001; ns, no significance.
    Sw1353, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 745 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/sw+1353/SW+1353/pmc13214264-49-0-2
    Average 96 stars, based on 745 article reviews
    sw1353 - by Bioz Stars, 2026-09
    96/100 stars

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    1) Product Images from "Mitochondrial dysfunction activates ADAMTS-5 expression via mt-dsRNA-PKR-Spi-1 axis in osteoarthritic chondrocytes"

    Article Title: Mitochondrial dysfunction activates ADAMTS-5 expression via mt-dsRNA-PKR-Spi-1 axis in osteoarthritic chondrocytes

    Journal: iScience

    doi: 10.1016/j.isci.2026.115980

    Mitochondrial dysfunction activates ADAMTS-5 expression in chondrocytes (A–D) The impact of inhibiting mitochondrial respiratory chain with oligo A (30 μg/mL) on mitochondrial membrane potential (A, n = 6), ROS generation (B, n = 6), cell survival (C, n = 6), and the mRNA expression of IFNB1 and ISGs (D, n = 6) in the SW1353 cell line. Scale bars in (B) are 20 μm. (E) Immunoblotting was used to analyze the phosphorylation of PKR and eIF2α following oligo A treatment in the SW1353 cell line, with quantification displayed on the right ( n = 6). (F and G) RT-qPCR and immunoblotting were conducted to assess the expression of ADAMTS-5 mRNA (F, n = 6) and protein (G) in response to oligo A treatment in the SW1353 cell line, with immunoblotting quantification shown on the right ( n = 6). Data (mean ± std) was representative of three independent experiments. An unpaired student’s t test was conducted to compare two groups. ∗, p < 0.05, ∗, p < 0.01, ∗∗∗, p < 0.001; ns, no significance.
    Figure Legend Snippet: Mitochondrial dysfunction activates ADAMTS-5 expression in chondrocytes (A–D) The impact of inhibiting mitochondrial respiratory chain with oligo A (30 μg/mL) on mitochondrial membrane potential (A, n = 6), ROS generation (B, n = 6), cell survival (C, n = 6), and the mRNA expression of IFNB1 and ISGs (D, n = 6) in the SW1353 cell line. Scale bars in (B) are 20 μm. (E) Immunoblotting was used to analyze the phosphorylation of PKR and eIF2α following oligo A treatment in the SW1353 cell line, with quantification displayed on the right ( n = 6). (F and G) RT-qPCR and immunoblotting were conducted to assess the expression of ADAMTS-5 mRNA (F, n = 6) and protein (G) in response to oligo A treatment in the SW1353 cell line, with immunoblotting quantification shown on the right ( n = 6). Data (mean ± std) was representative of three independent experiments. An unpaired student’s t test was conducted to compare two groups. ∗, p < 0.05, ∗, p < 0.01, ∗∗∗, p < 0.001; ns, no significance.

    Techniques Used: Expressing, Membrane, Western Blot, Phospho-proteomics, Quantitative RT-PCR

    Mitochondrial dysfunction activates ADAMTS-5 expression via PKR in chondrocytes (A) Analysis of PKR overexpression introduced via a lentivirus vector carrying the PKR coding sequence in the SW1353 cell line using immunoblotting. (B) Examination of IFNB1 and ISG mRNA levels following PKR overexpression ( n = 6). (C and D) Evaluation of ADAMTS-5 mRNA (C, n = 6) and protein (D) levels in response to PKR overexpression in the SW1353 cell line through RT-qPCR and immunoblotting. The quantification of immunoblotting results is displayed on the right ( n = 6). (E and F) Analysis of PKR mRNA (E, n = 6) and protein (F) levels following siRNA-mediated PKR knockdown in the SW1353 cell line using RT-qPCR and immunoblotting. The quantification of immunoblotting results is shown on the right ( n = 6). (G and H) Assessment of cell viability (G, n = 6), and IFNB1 and ISG mRNA levels (H, n = 6) in response to oligo A treatment and/or PKR knockdown in the SW1353 cell line. (I and J) Evaluation of ADAMTS-5 mRNA (I, n = 6) and protein (J) levels in response to oligo A treatment, PKR knockdown and/or recombinant human PKR protein treatment in the SW1353 cell line using RT-qPCR and immunoblotting. The quantification of immunoblotting results is displayed on the right ( n = 6). (K) Impact of mutating one of the two critical phosphorylation residues (T446A or T451A) of PKR on cell viability ( n = 6). (L and M) Analysis of ADAMTS-5 mRNA (L, n = 6) and protein (M) levels in response to the mutation of one of the two key phosphorylation residues (T446A or T451A) of PKR in the SW1353 cell line using RT-qPCR and immunoblotting. The quantification of immunoblotting results is shown on the right ( n = 6). Data (mean ± std) was representative of three independent experiments. To compare two groups, an unpaired student’s t test was conducted in (B, C, D, E, F, and H). For analyses involving multiple groups, one-way (K, L, and M) and two-way (G, I, and J) ANOVA were employed. ∗, p < 0.05, ∗, p < 0.01, ∗∗∗, p < 0.001; ns, no significance.
    Figure Legend Snippet: Mitochondrial dysfunction activates ADAMTS-5 expression via PKR in chondrocytes (A) Analysis of PKR overexpression introduced via a lentivirus vector carrying the PKR coding sequence in the SW1353 cell line using immunoblotting. (B) Examination of IFNB1 and ISG mRNA levels following PKR overexpression ( n = 6). (C and D) Evaluation of ADAMTS-5 mRNA (C, n = 6) and protein (D) levels in response to PKR overexpression in the SW1353 cell line through RT-qPCR and immunoblotting. The quantification of immunoblotting results is displayed on the right ( n = 6). (E and F) Analysis of PKR mRNA (E, n = 6) and protein (F) levels following siRNA-mediated PKR knockdown in the SW1353 cell line using RT-qPCR and immunoblotting. The quantification of immunoblotting results is shown on the right ( n = 6). (G and H) Assessment of cell viability (G, n = 6), and IFNB1 and ISG mRNA levels (H, n = 6) in response to oligo A treatment and/or PKR knockdown in the SW1353 cell line. (I and J) Evaluation of ADAMTS-5 mRNA (I, n = 6) and protein (J) levels in response to oligo A treatment, PKR knockdown and/or recombinant human PKR protein treatment in the SW1353 cell line using RT-qPCR and immunoblotting. The quantification of immunoblotting results is displayed on the right ( n = 6). (K) Impact of mutating one of the two critical phosphorylation residues (T446A or T451A) of PKR on cell viability ( n = 6). (L and M) Analysis of ADAMTS-5 mRNA (L, n = 6) and protein (M) levels in response to the mutation of one of the two key phosphorylation residues (T446A or T451A) of PKR in the SW1353 cell line using RT-qPCR and immunoblotting. The quantification of immunoblotting results is shown on the right ( n = 6). Data (mean ± std) was representative of three independent experiments. To compare two groups, an unpaired student’s t test was conducted in (B, C, D, E, F, and H). For analyses involving multiple groups, one-way (K, L, and M) and two-way (G, I, and J) ANOVA were employed. ∗, p < 0.05, ∗, p < 0.01, ∗∗∗, p < 0.001; ns, no significance.

    Techniques Used: Expressing, Over Expression, Plasmid Preparation, Sequencing, Western Blot, Quantitative RT-PCR, Knockdown, Recombinant, Phospho-proteomics, Mutagenesis

    Mitochondrial dysfunction induces the cytosolic efflux of mt-dsRNAs to activate PKR (A) The interaction between PKR and mtRNA following oligo A treatment was assessed using PKR fCLIP-qPCR analysis ( n = 6). (B) The interaction of PKR with mtRNA was analyzed in the presence of wild-type (WT) and dsRBD-mut PKR through fCLIP-qPCR analysis ( n = 6). (C) The levels of mtRNA in the cytoplasm of the SW1353 cell line were measured in response to oligo A treatment using RT-qPCR ( n = 6). (D) The interaction between PKR and mtRNA after 2-CM (50 μM) treatment was evaluated using PKR fCLIP-qPCR analysis ( n = 6). (E–I) The mitochondrial membrane potential (E, n = 6), ROS production (F, n = 6), cell viability (G, n = 6), phosphorylation of PKR and eIF2α (H), and the expression of IFNB1 and ISGs mRNA (I, n = 6) were examined in response to oligo A and 2-CM treatment in the SW1353 cell line. The scale bars in (F) are 20 μm. The quantification of the western blot is displayed to the right of the blot images ( n = 6). (J and K) RT-qPCR and immunoblotting were used to analyze the expression of ADAMTS-5 mRNA (J, n = 6) and protein (K) in response to oligo A and 2-CM treatment in the SW1353 cell line. The quantification of immunoblotting is shown on the right ( n = 6). Data (mean ± std) was representative of three independent experiments. An unpaired student’s t test was conducted to compare two groups. ∗, p < 0.05, ∗, p < 0.01, ∗∗∗, p < 0.001; ns, no significance.
    Figure Legend Snippet: Mitochondrial dysfunction induces the cytosolic efflux of mt-dsRNAs to activate PKR (A) The interaction between PKR and mtRNA following oligo A treatment was assessed using PKR fCLIP-qPCR analysis ( n = 6). (B) The interaction of PKR with mtRNA was analyzed in the presence of wild-type (WT) and dsRBD-mut PKR through fCLIP-qPCR analysis ( n = 6). (C) The levels of mtRNA in the cytoplasm of the SW1353 cell line were measured in response to oligo A treatment using RT-qPCR ( n = 6). (D) The interaction between PKR and mtRNA after 2-CM (50 μM) treatment was evaluated using PKR fCLIP-qPCR analysis ( n = 6). (E–I) The mitochondrial membrane potential (E, n = 6), ROS production (F, n = 6), cell viability (G, n = 6), phosphorylation of PKR and eIF2α (H), and the expression of IFNB1 and ISGs mRNA (I, n = 6) were examined in response to oligo A and 2-CM treatment in the SW1353 cell line. The scale bars in (F) are 20 μm. The quantification of the western blot is displayed to the right of the blot images ( n = 6). (J and K) RT-qPCR and immunoblotting were used to analyze the expression of ADAMTS-5 mRNA (J, n = 6) and protein (K) in response to oligo A and 2-CM treatment in the SW1353 cell line. The quantification of immunoblotting is shown on the right ( n = 6). Data (mean ± std) was representative of three independent experiments. An unpaired student’s t test was conducted to compare two groups. ∗, p < 0.05, ∗, p < 0.01, ∗∗∗, p < 0.001; ns, no significance.

    Techniques Used: Quantitative RT-PCR, Membrane, Phospho-proteomics, Expressing, Western Blot

    PKR activated by mt-dsRNAs enhances the translocation of Spi-1 to nuclear and its binding on ADAMTS-5 promoter (A and B) The expression of Spi-1 mRNA (A, n = 6) and protein (B) in the SW1353 cell line was analyzed using RT-qPCR and immunoblotting following oligo A treatment. The quantification of the immunoblotting results is displayed on the right ( n = 6). (C) Immunoblotting was used to assess Spi-1 protein levels in the nucleus of the SW1353 cell line after oligo A treatment, with quantification shown on the right ( n = 6). (D) The binding of Spi-1 to the ADAMTS-5 promoter in response to oligo A treatment was evaluated using ChIP, with quantification on the right ( n = 6). (E) The nuclear expression of Spi-1 protein in the SW1353 cell line was examined through immunoblotting after treatment with oligo A and 2-CM, with quantification on the right ( n = 6). (F) ChIP was used to investigate Spi-1 binding to the ADAMTS-5 promoter following oligo A and 2-CM treatment, with quantification on the right ( n = 6). (G) Immunoblotting analysis was conducted to determine Spi-1 protein levels in the nucleus of the SW1353 cell line after oligo A treatment and PKR knockdown, with quantification on the right ( n = 6). (H) ChIP analysis was performed to assess Spi-1 binding to the ADAMTS-5 promoter in response to oligo A treatment and PKR knockdown, with quantification on the right ( n = 6). (I and J) The expression of ADAMTS-5 mRNA (I, n = 6) and protein (J) in the SW1353 cell line was analyzed using RT-qPCR and immunoblotting following oligo A treatment, Spi-1 knockdown and/or recombinant human Spi-1 treatment, with quantification on the right ( n = 6). Data (mean ± std) was representative of three independent experiments. To compare two groups, an unpaired Student’s t test was conducted in (A–H). For analyses involving multiple groups, two-way (I and J) ANOVA were employed. ∗, p < 0.05, ∗, p < 0.01, ∗∗∗, p < 0.001; ns, no significance.
    Figure Legend Snippet: PKR activated by mt-dsRNAs enhances the translocation of Spi-1 to nuclear and its binding on ADAMTS-5 promoter (A and B) The expression of Spi-1 mRNA (A, n = 6) and protein (B) in the SW1353 cell line was analyzed using RT-qPCR and immunoblotting following oligo A treatment. The quantification of the immunoblotting results is displayed on the right ( n = 6). (C) Immunoblotting was used to assess Spi-1 protein levels in the nucleus of the SW1353 cell line after oligo A treatment, with quantification shown on the right ( n = 6). (D) The binding of Spi-1 to the ADAMTS-5 promoter in response to oligo A treatment was evaluated using ChIP, with quantification on the right ( n = 6). (E) The nuclear expression of Spi-1 protein in the SW1353 cell line was examined through immunoblotting after treatment with oligo A and 2-CM, with quantification on the right ( n = 6). (F) ChIP was used to investigate Spi-1 binding to the ADAMTS-5 promoter following oligo A and 2-CM treatment, with quantification on the right ( n = 6). (G) Immunoblotting analysis was conducted to determine Spi-1 protein levels in the nucleus of the SW1353 cell line after oligo A treatment and PKR knockdown, with quantification on the right ( n = 6). (H) ChIP analysis was performed to assess Spi-1 binding to the ADAMTS-5 promoter in response to oligo A treatment and PKR knockdown, with quantification on the right ( n = 6). (I and J) The expression of ADAMTS-5 mRNA (I, n = 6) and protein (J) in the SW1353 cell line was analyzed using RT-qPCR and immunoblotting following oligo A treatment, Spi-1 knockdown and/or recombinant human Spi-1 treatment, with quantification on the right ( n = 6). Data (mean ± std) was representative of three independent experiments. To compare two groups, an unpaired Student’s t test was conducted in (A–H). For analyses involving multiple groups, two-way (I and J) ANOVA were employed. ∗, p < 0.05, ∗, p < 0.01, ∗∗∗, p < 0.001; ns, no significance.

    Techniques Used: Translocation Assay, Binding Assay, Expressing, Quantitative RT-PCR, Western Blot, Knockdown, Recombinant

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    In vitro tube formation ability by chondrosarcoma cells. ( A ) Representative images of HUVEC (upper panels) or Sarc cells (lower panels) seeded onto GFR-Matrigel and allowed to form vascular tubules in vitro in the absence of serum (CTRL) or in the presence of FBS. Images acquired using an inverted phase-contrast microscope (Axiovert 200) and Axiovision 4.8 software (Zeiss). Scale bar, 100 μm or 50 μm in panels with higher magnification. Periodic acid—Schiff staining performed on HUVECs and Sarc cells (panels on the right) effectively shows that the newly formed tubes are rich in PAS-positive glycogen. ( B ) Representative images of <t>SW1353</t> cell line and patient-derived ChS cells (ChS-1, ChS-2, and ChS-3), seeded onto GFR-Matrigel and allowed to form vascular-like tubules. Images acquired using an inverted phase-contrast microscope (Axiovert 200) and Axiovision 4.8 software (Zeiss). Scale bar, 500 μm.
    Sw1353 Chondrosarcoma Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Mitochondrial dysfunction activates ADAMTS-5 expression in chondrocytes (A–D) The impact of inhibiting mitochondrial respiratory chain with oligo A (30 μg/mL) on mitochondrial membrane potential (A, n = 6), ROS generation (B, n = 6), cell survival (C, n = 6), and the mRNA expression of IFNB1 and ISGs (D, n = 6) in the SW1353 cell line. Scale bars in (B) are 20 μm. (E) Immunoblotting was used to analyze the phosphorylation of PKR and eIF2α following oligo A treatment in the SW1353 cell line, with quantification displayed on the right ( n = 6). (F and G) RT-qPCR and immunoblotting were conducted to assess the expression of ADAMTS-5 mRNA (F, n = 6) and protein (G) in response to oligo A treatment in the SW1353 cell line, with immunoblotting quantification shown on the right ( n = 6). Data (mean ± std) was representative of three independent experiments. An unpaired student’s t test was conducted to compare two groups. ∗, p < 0.05, ∗, p < 0.01, ∗∗∗, p < 0.001; ns, no significance.

    Journal: iScience

    Article Title: Mitochondrial dysfunction activates ADAMTS-5 expression via mt-dsRNA-PKR-Spi-1 axis in osteoarthritic chondrocytes

    doi: 10.1016/j.isci.2026.115980

    Figure Lengend Snippet: Mitochondrial dysfunction activates ADAMTS-5 expression in chondrocytes (A–D) The impact of inhibiting mitochondrial respiratory chain with oligo A (30 μg/mL) on mitochondrial membrane potential (A, n = 6), ROS generation (B, n = 6), cell survival (C, n = 6), and the mRNA expression of IFNB1 and ISGs (D, n = 6) in the SW1353 cell line. Scale bars in (B) are 20 μm. (E) Immunoblotting was used to analyze the phosphorylation of PKR and eIF2α following oligo A treatment in the SW1353 cell line, with quantification displayed on the right ( n = 6). (F and G) RT-qPCR and immunoblotting were conducted to assess the expression of ADAMTS-5 mRNA (F, n = 6) and protein (G) in response to oligo A treatment in the SW1353 cell line, with immunoblotting quantification shown on the right ( n = 6). Data (mean ± std) was representative of three independent experiments. An unpaired student’s t test was conducted to compare two groups. ∗, p < 0.05, ∗, p < 0.01, ∗∗∗, p < 0.001; ns, no significance.

    Article Snippet: SW1353 , ATCC , HTB-94.

    Techniques: Expressing, Membrane, Western Blot, Phospho-proteomics, Quantitative RT-PCR

    Mitochondrial dysfunction activates ADAMTS-5 expression via PKR in chondrocytes (A) Analysis of PKR overexpression introduced via a lentivirus vector carrying the PKR coding sequence in the SW1353 cell line using immunoblotting. (B) Examination of IFNB1 and ISG mRNA levels following PKR overexpression ( n = 6). (C and D) Evaluation of ADAMTS-5 mRNA (C, n = 6) and protein (D) levels in response to PKR overexpression in the SW1353 cell line through RT-qPCR and immunoblotting. The quantification of immunoblotting results is displayed on the right ( n = 6). (E and F) Analysis of PKR mRNA (E, n = 6) and protein (F) levels following siRNA-mediated PKR knockdown in the SW1353 cell line using RT-qPCR and immunoblotting. The quantification of immunoblotting results is shown on the right ( n = 6). (G and H) Assessment of cell viability (G, n = 6), and IFNB1 and ISG mRNA levels (H, n = 6) in response to oligo A treatment and/or PKR knockdown in the SW1353 cell line. (I and J) Evaluation of ADAMTS-5 mRNA (I, n = 6) and protein (J) levels in response to oligo A treatment, PKR knockdown and/or recombinant human PKR protein treatment in the SW1353 cell line using RT-qPCR and immunoblotting. The quantification of immunoblotting results is displayed on the right ( n = 6). (K) Impact of mutating one of the two critical phosphorylation residues (T446A or T451A) of PKR on cell viability ( n = 6). (L and M) Analysis of ADAMTS-5 mRNA (L, n = 6) and protein (M) levels in response to the mutation of one of the two key phosphorylation residues (T446A or T451A) of PKR in the SW1353 cell line using RT-qPCR and immunoblotting. The quantification of immunoblotting results is shown on the right ( n = 6). Data (mean ± std) was representative of three independent experiments. To compare two groups, an unpaired student’s t test was conducted in (B, C, D, E, F, and H). For analyses involving multiple groups, one-way (K, L, and M) and two-way (G, I, and J) ANOVA were employed. ∗, p < 0.05, ∗, p < 0.01, ∗∗∗, p < 0.001; ns, no significance.

    Journal: iScience

    Article Title: Mitochondrial dysfunction activates ADAMTS-5 expression via mt-dsRNA-PKR-Spi-1 axis in osteoarthritic chondrocytes

    doi: 10.1016/j.isci.2026.115980

    Figure Lengend Snippet: Mitochondrial dysfunction activates ADAMTS-5 expression via PKR in chondrocytes (A) Analysis of PKR overexpression introduced via a lentivirus vector carrying the PKR coding sequence in the SW1353 cell line using immunoblotting. (B) Examination of IFNB1 and ISG mRNA levels following PKR overexpression ( n = 6). (C and D) Evaluation of ADAMTS-5 mRNA (C, n = 6) and protein (D) levels in response to PKR overexpression in the SW1353 cell line through RT-qPCR and immunoblotting. The quantification of immunoblotting results is displayed on the right ( n = 6). (E and F) Analysis of PKR mRNA (E, n = 6) and protein (F) levels following siRNA-mediated PKR knockdown in the SW1353 cell line using RT-qPCR and immunoblotting. The quantification of immunoblotting results is shown on the right ( n = 6). (G and H) Assessment of cell viability (G, n = 6), and IFNB1 and ISG mRNA levels (H, n = 6) in response to oligo A treatment and/or PKR knockdown in the SW1353 cell line. (I and J) Evaluation of ADAMTS-5 mRNA (I, n = 6) and protein (J) levels in response to oligo A treatment, PKR knockdown and/or recombinant human PKR protein treatment in the SW1353 cell line using RT-qPCR and immunoblotting. The quantification of immunoblotting results is displayed on the right ( n = 6). (K) Impact of mutating one of the two critical phosphorylation residues (T446A or T451A) of PKR on cell viability ( n = 6). (L and M) Analysis of ADAMTS-5 mRNA (L, n = 6) and protein (M) levels in response to the mutation of one of the two key phosphorylation residues (T446A or T451A) of PKR in the SW1353 cell line using RT-qPCR and immunoblotting. The quantification of immunoblotting results is shown on the right ( n = 6). Data (mean ± std) was representative of three independent experiments. To compare two groups, an unpaired student’s t test was conducted in (B, C, D, E, F, and H). For analyses involving multiple groups, one-way (K, L, and M) and two-way (G, I, and J) ANOVA were employed. ∗, p < 0.05, ∗, p < 0.01, ∗∗∗, p < 0.001; ns, no significance.

    Article Snippet: SW1353 , ATCC , HTB-94.

    Techniques: Expressing, Over Expression, Plasmid Preparation, Sequencing, Western Blot, Quantitative RT-PCR, Knockdown, Recombinant, Phospho-proteomics, Mutagenesis

    Mitochondrial dysfunction induces the cytosolic efflux of mt-dsRNAs to activate PKR (A) The interaction between PKR and mtRNA following oligo A treatment was assessed using PKR fCLIP-qPCR analysis ( n = 6). (B) The interaction of PKR with mtRNA was analyzed in the presence of wild-type (WT) and dsRBD-mut PKR through fCLIP-qPCR analysis ( n = 6). (C) The levels of mtRNA in the cytoplasm of the SW1353 cell line were measured in response to oligo A treatment using RT-qPCR ( n = 6). (D) The interaction between PKR and mtRNA after 2-CM (50 μM) treatment was evaluated using PKR fCLIP-qPCR analysis ( n = 6). (E–I) The mitochondrial membrane potential (E, n = 6), ROS production (F, n = 6), cell viability (G, n = 6), phosphorylation of PKR and eIF2α (H), and the expression of IFNB1 and ISGs mRNA (I, n = 6) were examined in response to oligo A and 2-CM treatment in the SW1353 cell line. The scale bars in (F) are 20 μm. The quantification of the western blot is displayed to the right of the blot images ( n = 6). (J and K) RT-qPCR and immunoblotting were used to analyze the expression of ADAMTS-5 mRNA (J, n = 6) and protein (K) in response to oligo A and 2-CM treatment in the SW1353 cell line. The quantification of immunoblotting is shown on the right ( n = 6). Data (mean ± std) was representative of three independent experiments. An unpaired student’s t test was conducted to compare two groups. ∗, p < 0.05, ∗, p < 0.01, ∗∗∗, p < 0.001; ns, no significance.

    Journal: iScience

    Article Title: Mitochondrial dysfunction activates ADAMTS-5 expression via mt-dsRNA-PKR-Spi-1 axis in osteoarthritic chondrocytes

    doi: 10.1016/j.isci.2026.115980

    Figure Lengend Snippet: Mitochondrial dysfunction induces the cytosolic efflux of mt-dsRNAs to activate PKR (A) The interaction between PKR and mtRNA following oligo A treatment was assessed using PKR fCLIP-qPCR analysis ( n = 6). (B) The interaction of PKR with mtRNA was analyzed in the presence of wild-type (WT) and dsRBD-mut PKR through fCLIP-qPCR analysis ( n = 6). (C) The levels of mtRNA in the cytoplasm of the SW1353 cell line were measured in response to oligo A treatment using RT-qPCR ( n = 6). (D) The interaction between PKR and mtRNA after 2-CM (50 μM) treatment was evaluated using PKR fCLIP-qPCR analysis ( n = 6). (E–I) The mitochondrial membrane potential (E, n = 6), ROS production (F, n = 6), cell viability (G, n = 6), phosphorylation of PKR and eIF2α (H), and the expression of IFNB1 and ISGs mRNA (I, n = 6) were examined in response to oligo A and 2-CM treatment in the SW1353 cell line. The scale bars in (F) are 20 μm. The quantification of the western blot is displayed to the right of the blot images ( n = 6). (J and K) RT-qPCR and immunoblotting were used to analyze the expression of ADAMTS-5 mRNA (J, n = 6) and protein (K) in response to oligo A and 2-CM treatment in the SW1353 cell line. The quantification of immunoblotting is shown on the right ( n = 6). Data (mean ± std) was representative of three independent experiments. An unpaired student’s t test was conducted to compare two groups. ∗, p < 0.05, ∗, p < 0.01, ∗∗∗, p < 0.001; ns, no significance.

    Article Snippet: SW1353 , ATCC , HTB-94.

    Techniques: Quantitative RT-PCR, Membrane, Phospho-proteomics, Expressing, Western Blot

    PKR activated by mt-dsRNAs enhances the translocation of Spi-1 to nuclear and its binding on ADAMTS-5 promoter (A and B) The expression of Spi-1 mRNA (A, n = 6) and protein (B) in the SW1353 cell line was analyzed using RT-qPCR and immunoblotting following oligo A treatment. The quantification of the immunoblotting results is displayed on the right ( n = 6). (C) Immunoblotting was used to assess Spi-1 protein levels in the nucleus of the SW1353 cell line after oligo A treatment, with quantification shown on the right ( n = 6). (D) The binding of Spi-1 to the ADAMTS-5 promoter in response to oligo A treatment was evaluated using ChIP, with quantification on the right ( n = 6). (E) The nuclear expression of Spi-1 protein in the SW1353 cell line was examined through immunoblotting after treatment with oligo A and 2-CM, with quantification on the right ( n = 6). (F) ChIP was used to investigate Spi-1 binding to the ADAMTS-5 promoter following oligo A and 2-CM treatment, with quantification on the right ( n = 6). (G) Immunoblotting analysis was conducted to determine Spi-1 protein levels in the nucleus of the SW1353 cell line after oligo A treatment and PKR knockdown, with quantification on the right ( n = 6). (H) ChIP analysis was performed to assess Spi-1 binding to the ADAMTS-5 promoter in response to oligo A treatment and PKR knockdown, with quantification on the right ( n = 6). (I and J) The expression of ADAMTS-5 mRNA (I, n = 6) and protein (J) in the SW1353 cell line was analyzed using RT-qPCR and immunoblotting following oligo A treatment, Spi-1 knockdown and/or recombinant human Spi-1 treatment, with quantification on the right ( n = 6). Data (mean ± std) was representative of three independent experiments. To compare two groups, an unpaired Student’s t test was conducted in (A–H). For analyses involving multiple groups, two-way (I and J) ANOVA were employed. ∗, p < 0.05, ∗, p < 0.01, ∗∗∗, p < 0.001; ns, no significance.

    Journal: iScience

    Article Title: Mitochondrial dysfunction activates ADAMTS-5 expression via mt-dsRNA-PKR-Spi-1 axis in osteoarthritic chondrocytes

    doi: 10.1016/j.isci.2026.115980

    Figure Lengend Snippet: PKR activated by mt-dsRNAs enhances the translocation of Spi-1 to nuclear and its binding on ADAMTS-5 promoter (A and B) The expression of Spi-1 mRNA (A, n = 6) and protein (B) in the SW1353 cell line was analyzed using RT-qPCR and immunoblotting following oligo A treatment. The quantification of the immunoblotting results is displayed on the right ( n = 6). (C) Immunoblotting was used to assess Spi-1 protein levels in the nucleus of the SW1353 cell line after oligo A treatment, with quantification shown on the right ( n = 6). (D) The binding of Spi-1 to the ADAMTS-5 promoter in response to oligo A treatment was evaluated using ChIP, with quantification on the right ( n = 6). (E) The nuclear expression of Spi-1 protein in the SW1353 cell line was examined through immunoblotting after treatment with oligo A and 2-CM, with quantification on the right ( n = 6). (F) ChIP was used to investigate Spi-1 binding to the ADAMTS-5 promoter following oligo A and 2-CM treatment, with quantification on the right ( n = 6). (G) Immunoblotting analysis was conducted to determine Spi-1 protein levels in the nucleus of the SW1353 cell line after oligo A treatment and PKR knockdown, with quantification on the right ( n = 6). (H) ChIP analysis was performed to assess Spi-1 binding to the ADAMTS-5 promoter in response to oligo A treatment and PKR knockdown, with quantification on the right ( n = 6). (I and J) The expression of ADAMTS-5 mRNA (I, n = 6) and protein (J) in the SW1353 cell line was analyzed using RT-qPCR and immunoblotting following oligo A treatment, Spi-1 knockdown and/or recombinant human Spi-1 treatment, with quantification on the right ( n = 6). Data (mean ± std) was representative of three independent experiments. To compare two groups, an unpaired Student’s t test was conducted in (A–H). For analyses involving multiple groups, two-way (I and J) ANOVA were employed. ∗, p < 0.05, ∗, p < 0.01, ∗∗∗, p < 0.001; ns, no significance.

    Article Snippet: SW1353 , ATCC , HTB-94.

    Techniques: Translocation Assay, Binding Assay, Expressing, Quantitative RT-PCR, Western Blot, Knockdown, Recombinant

    RT-qPCR Validation of Key Genes in Chondrosarcoma Cell Lines. (A-D) Relative expression levels of LINC00984, TXNDC2, MPL28, and RP11-474N24.6 in SW1353, JJ012, and OUMS-27 chondrosarcoma cell lines compared to normal C28/I2 chondrocytes. Data represent mean ± SD from three independent experiments. *** P < 0.001, ** P < 0.01 vs. control group

    Journal: Discover Oncology

    Article Title: Mendelian randomization analysis of causal relationships between chondrosarcoma and angiogenesis related genes

    doi: 10.1007/s12672-026-04621-0

    Figure Lengend Snippet: RT-qPCR Validation of Key Genes in Chondrosarcoma Cell Lines. (A-D) Relative expression levels of LINC00984, TXNDC2, MPL28, and RP11-474N24.6 in SW1353, JJ012, and OUMS-27 chondrosarcoma cell lines compared to normal C28/I2 chondrocytes. Data represent mean ± SD from three independent experiments. *** P < 0.001, ** P < 0.01 vs. control group

    Article Snippet: Human chondrosarcoma cell lines SW1353, JJ012, and OUMS-27, along with normal human chondrocyte cell line C28/I2 (control), were obtained from ATCC and cultured in DMEM supplemented with 10% fetal bovine serum, 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C with 5% CO2.

    Techniques: Quantitative RT-PCR, Biomarker Discovery, Expressing, Control

    ( a ) The LGALS3 full-length promoter (−2638/+52) and the deletions (−1925/+52), (−1442/+52), (−535/+52), (−428/+52), (−328/+52), (−228/+52), (−117/+52), (−97/+52), (−77/+52), (−37/+52), (−17/+52) are shown in a diagram drawn to scale. Positions are numbered according to the distance to the transcription start site (TSS), where negative means upstream and positive means downstream of the TSS. ( b ) Relative activity of LGALS3 promoter sequence deletions in SW1353 cells. The activity of these deletions is shown as an x-fold increase relative to cells transfected with the promoter-less vector pGL4.20 (Basic). The graph shows the results’ mean ± SD of at least four biological replicates, each including three technical replicates, with error bars representing the standard deviation. * p < 0.05.

    Journal: Scientific Reports

    Article Title: SOX9 represses the human galectin-3 promoter in SW1353 cells: potential implications for osteoarthritis

    doi: 10.1038/s41598-026-50507-0

    Figure Lengend Snippet: ( a ) The LGALS3 full-length promoter (−2638/+52) and the deletions (−1925/+52), (−1442/+52), (−535/+52), (−428/+52), (−328/+52), (−228/+52), (−117/+52), (−97/+52), (−77/+52), (−37/+52), (−17/+52) are shown in a diagram drawn to scale. Positions are numbered according to the distance to the transcription start site (TSS), where negative means upstream and positive means downstream of the TSS. ( b ) Relative activity of LGALS3 promoter sequence deletions in SW1353 cells. The activity of these deletions is shown as an x-fold increase relative to cells transfected with the promoter-less vector pGL4.20 (Basic). The graph shows the results’ mean ± SD of at least four biological replicates, each including three technical replicates, with error bars representing the standard deviation. * p < 0.05.

    Article Snippet: SW1353 human chondrosarcoma cells (ATCC HTB-94TM, American Type Culture Collection (ATCC)) were cultured at 37 °C in a humidified atmosphere with 5% CO 2 in Dulbecco’s Modified Eagles Medium (DMEM), containing 10% FCS and 1% penicillin/streptomycin/L-glutamine.

    Techniques: Activity Assay, Sequencing, Transfection, Plasmid Preparation, Standard Deviation

    Relative activity of the LGALS3 full-length promoter (−2638/+52) in SW1353 cells when co-transfected with 30 ng of different SOX-containing plasmids (pcDNA3.1(+)-SOX2, -SOX4, -SOX5, -SOX6 and -SOX9). The relative activity of the promoter in the different conditions is calculated as a ratio to the promoter activity of the cells transfected with the vector pcDNA3.1(+) lacking the respective SOX. The graph shows results’ mean relative activity ± SD from four technical replicates, each including three biological replicates, with error bars representing the standard deviation. * p < 0.05.

    Journal: Scientific Reports

    Article Title: SOX9 represses the human galectin-3 promoter in SW1353 cells: potential implications for osteoarthritis

    doi: 10.1038/s41598-026-50507-0

    Figure Lengend Snippet: Relative activity of the LGALS3 full-length promoter (−2638/+52) in SW1353 cells when co-transfected with 30 ng of different SOX-containing plasmids (pcDNA3.1(+)-SOX2, -SOX4, -SOX5, -SOX6 and -SOX9). The relative activity of the promoter in the different conditions is calculated as a ratio to the promoter activity of the cells transfected with the vector pcDNA3.1(+) lacking the respective SOX. The graph shows results’ mean relative activity ± SD from four technical replicates, each including three biological replicates, with error bars representing the standard deviation. * p < 0.05.

    Article Snippet: SW1353 human chondrosarcoma cells (ATCC HTB-94TM, American Type Culture Collection (ATCC)) were cultured at 37 °C in a humidified atmosphere with 5% CO 2 in Dulbecco’s Modified Eagles Medium (DMEM), containing 10% FCS and 1% penicillin/streptomycin/L-glutamine.

    Techniques: Activity Assay, Transfection, Plasmid Preparation, Standard Deviation

    Relative activity of the LGALS3 full-length promoter (−2638/+52) in SW1353 cells when titrated with different amounts of SOX-containing plasmids. ( a ) pcDNA3.1(+)-SOX2, ( b ) pcDNA3.1(+)-SOX4, ( c ) pcDNA3.1(+)-SOX5, ( d ) pcDNA3.1(+)-SOX6, ( e ) pcDNA3.1(+)-SOX9. The amounts of DNA used were: 0.1 ng, 1 ng, 10 ng, 100 ng. For each condition, a negative control without the SOX-encoding plasmid was included. The x-axis is plotted on a logarithmic scale. The relative activity of the promoter in the different conditions is calculated as a ratio to the promoter activity of the cells transfected with the vector pcDNA3.1(+) lacking the respective SOX. The graph shows results’ mean relative activity ± SD from three biological replicates, including three technical replicates, with error bars representing the standard deviation. * p < 0.05.

    Journal: Scientific Reports

    Article Title: SOX9 represses the human galectin-3 promoter in SW1353 cells: potential implications for osteoarthritis

    doi: 10.1038/s41598-026-50507-0

    Figure Lengend Snippet: Relative activity of the LGALS3 full-length promoter (−2638/+52) in SW1353 cells when titrated with different amounts of SOX-containing plasmids. ( a ) pcDNA3.1(+)-SOX2, ( b ) pcDNA3.1(+)-SOX4, ( c ) pcDNA3.1(+)-SOX5, ( d ) pcDNA3.1(+)-SOX6, ( e ) pcDNA3.1(+)-SOX9. The amounts of DNA used were: 0.1 ng, 1 ng, 10 ng, 100 ng. For each condition, a negative control without the SOX-encoding plasmid was included. The x-axis is plotted on a logarithmic scale. The relative activity of the promoter in the different conditions is calculated as a ratio to the promoter activity of the cells transfected with the vector pcDNA3.1(+) lacking the respective SOX. The graph shows results’ mean relative activity ± SD from three biological replicates, including three technical replicates, with error bars representing the standard deviation. * p < 0.05.

    Article Snippet: SW1353 human chondrosarcoma cells (ATCC HTB-94TM, American Type Culture Collection (ATCC)) were cultured at 37 °C in a humidified atmosphere with 5% CO 2 in Dulbecco’s Modified Eagles Medium (DMEM), containing 10% FCS and 1% penicillin/streptomycin/L-glutamine.

    Techniques: Activity Assay, Negative Control, Plasmid Preparation, Transfection, Standard Deviation

    Relative activity of LGALS3 promoter deletion variants in SW1353 cells when co-transfected for 24 h with SOX-protein-containing plasmids: ( a ) SOX2 or ( b ) SOX9. The graph shows results’ mean ± SD from at least three biological replicates, including three technical replicates, with error bars representing the standard deviation. The white bars represent the negative control (empty pcDNA3.1(+)). ( a ) The light grey bars represent SOX2. ( b ) The dark grey bars represent SOX9.

    Journal: Scientific Reports

    Article Title: SOX9 represses the human galectin-3 promoter in SW1353 cells: potential implications for osteoarthritis

    doi: 10.1038/s41598-026-50507-0

    Figure Lengend Snippet: Relative activity of LGALS3 promoter deletion variants in SW1353 cells when co-transfected for 24 h with SOX-protein-containing plasmids: ( a ) SOX2 or ( b ) SOX9. The graph shows results’ mean ± SD from at least three biological replicates, including three technical replicates, with error bars representing the standard deviation. The white bars represent the negative control (empty pcDNA3.1(+)). ( a ) The light grey bars represent SOX2. ( b ) The dark grey bars represent SOX9.

    Article Snippet: SW1353 human chondrosarcoma cells (ATCC HTB-94TM, American Type Culture Collection (ATCC)) were cultured at 37 °C in a humidified atmosphere with 5% CO 2 in Dulbecco’s Modified Eagles Medium (DMEM), containing 10% FCS and 1% penicillin/streptomycin/L-glutamine.

    Techniques: Activity Assay, Transfection, Standard Deviation, Negative Control

    Fold enrichment of specific promoter binding of HaloTag-SOX9 and hGal3p(−93/+49). ( a ) HaloChIP TM experiments were performed using SW1353 cells transfected with HaloTag-SOX9 or with the HaloTag-only vector (pHTN, control). DNA bound to SOX9 was isolated using the HaloCHIP™ system (Promega), and DNA was amplified via quantitative PCR. ADAMTS-4 is a known target of SOX9 and was used as a positive control. The graph shows mean ± SD from two biological replicates, with error bars representing the standard deviation. White bars represent the HaloTag-only control (pHTN). The dark grey bars represent HaloTag-SOX9. ( b ) The sequence stretch studied is shown, with a potential SOX9 binding site shown in a box. ( c ) A canonical SOX9 binding motif is shown.

    Journal: Scientific Reports

    Article Title: SOX9 represses the human galectin-3 promoter in SW1353 cells: potential implications for osteoarthritis

    doi: 10.1038/s41598-026-50507-0

    Figure Lengend Snippet: Fold enrichment of specific promoter binding of HaloTag-SOX9 and hGal3p(−93/+49). ( a ) HaloChIP TM experiments were performed using SW1353 cells transfected with HaloTag-SOX9 or with the HaloTag-only vector (pHTN, control). DNA bound to SOX9 was isolated using the HaloCHIP™ system (Promega), and DNA was amplified via quantitative PCR. ADAMTS-4 is a known target of SOX9 and was used as a positive control. The graph shows mean ± SD from two biological replicates, with error bars representing the standard deviation. White bars represent the HaloTag-only control (pHTN). The dark grey bars represent HaloTag-SOX9. ( b ) The sequence stretch studied is shown, with a potential SOX9 binding site shown in a box. ( c ) A canonical SOX9 binding motif is shown.

    Article Snippet: SW1353 human chondrosarcoma cells (ATCC HTB-94TM, American Type Culture Collection (ATCC)) were cultured at 37 °C in a humidified atmosphere with 5% CO 2 in Dulbecco’s Modified Eagles Medium (DMEM), containing 10% FCS and 1% penicillin/streptomycin/L-glutamine.

    Techniques: Binding Assay, Transfection, Plasmid Preparation, Control, Isolation, Amplification, Real-time Polymerase Chain Reaction, Positive Control, Standard Deviation, Sequencing

    Mutagenesis of the predicted SOX9 binding-site in the LGALS3 promoter. ( a ) The predicted SOX9 motif is shown within the wild-type sequence (WT); Mut shows the mutated motif. ( b ) Firefly luciferase activity (Firefly/ Renilla ratio) of the LGALS3 promoter construct (−97/+52) WT versus the LGALS3 promoter construct (−97/+52) carrying a mutation in the predicted SOX9 binding site in SW1353 cells, co-transfected with pcDNA (white bars) or pcDNA- SOX9 (grey bars). Data represent mean ± SD of three independent experiments performed in triplicate. Statistical significance is indicated as * p < 0.05, ** p < 0.01, *** p < 0.001.

    Journal: Scientific Reports

    Article Title: SOX9 represses the human galectin-3 promoter in SW1353 cells: potential implications for osteoarthritis

    doi: 10.1038/s41598-026-50507-0

    Figure Lengend Snippet: Mutagenesis of the predicted SOX9 binding-site in the LGALS3 promoter. ( a ) The predicted SOX9 motif is shown within the wild-type sequence (WT); Mut shows the mutated motif. ( b ) Firefly luciferase activity (Firefly/ Renilla ratio) of the LGALS3 promoter construct (−97/+52) WT versus the LGALS3 promoter construct (−97/+52) carrying a mutation in the predicted SOX9 binding site in SW1353 cells, co-transfected with pcDNA (white bars) or pcDNA- SOX9 (grey bars). Data represent mean ± SD of three independent experiments performed in triplicate. Statistical significance is indicated as * p < 0.05, ** p < 0.01, *** p < 0.001.

    Article Snippet: SW1353 human chondrosarcoma cells (ATCC HTB-94TM, American Type Culture Collection (ATCC)) were cultured at 37 °C in a humidified atmosphere with 5% CO 2 in Dulbecco’s Modified Eagles Medium (DMEM), containing 10% FCS and 1% penicillin/streptomycin/L-glutamine.

    Techniques: Mutagenesis, Binding Assay, Sequencing, Luciferase, Activity Assay, Construct, Transfection

    Effects of UE and UH on the production of IL-6, MMP-3, and Collagen 2α1 in MIA-induced SW1353 cells. Data are presented as mean ± SD ( n = 3). # indicates significant difference at p < 0.05; ## indicates significant difference at p < 0.01. Collagen 2α1, Collagen Type II Alpha 1; IL, interleukin; MIA, monosodium iodoacetate; MMP, matrix metalloproteinase; UE, Ulva extract; UH, Ulva hydrolysate

    Journal: Bioresources and Bioprocessing

    Article Title: Ulvan and Ulva oligosaccharides from Ulva sp. attenuate osteoarthritis in a high-fat diet and ligamentous meniscal injury-induced rat model

    doi: 10.1186/s40643-026-01012-9

    Figure Lengend Snippet: Effects of UE and UH on the production of IL-6, MMP-3, and Collagen 2α1 in MIA-induced SW1353 cells. Data are presented as mean ± SD ( n = 3). # indicates significant difference at p < 0.05; ## indicates significant difference at p < 0.01. Collagen 2α1, Collagen Type II Alpha 1; IL, interleukin; MIA, monosodium iodoacetate; MMP, matrix metalloproteinase; UE, Ulva extract; UH, Ulva hydrolysate

    Article Snippet: The human chondrosarcoma cell line SW1353 was purchased from the Bioresource Collection and Research Center of the Food Industry Research and Development Institute and the American Type Culture Collection (ATCC).

    Techniques:

    In vitro tube formation ability by chondrosarcoma cells. ( A ) Representative images of HUVEC (upper panels) or Sarc cells (lower panels) seeded onto GFR-Matrigel and allowed to form vascular tubules in vitro in the absence of serum (CTRL) or in the presence of FBS. Images acquired using an inverted phase-contrast microscope (Axiovert 200) and Axiovision 4.8 software (Zeiss). Scale bar, 100 μm or 50 μm in panels with higher magnification. Periodic acid—Schiff staining performed on HUVECs and Sarc cells (panels on the right) effectively shows that the newly formed tubes are rich in PAS-positive glycogen. ( B ) Representative images of SW1353 cell line and patient-derived ChS cells (ChS-1, ChS-2, and ChS-3), seeded onto GFR-Matrigel and allowed to form vascular-like tubules. Images acquired using an inverted phase-contrast microscope (Axiovert 200) and Axiovision 4.8 software (Zeiss). Scale bar, 500 μm.

    Journal: Cells

    Article Title: Vasculogenic Mimicry: A Potential Therapeutic Target for Chondrosarcoma Therapy

    doi: 10.3390/cells15050392

    Figure Lengend Snippet: In vitro tube formation ability by chondrosarcoma cells. ( A ) Representative images of HUVEC (upper panels) or Sarc cells (lower panels) seeded onto GFR-Matrigel and allowed to form vascular tubules in vitro in the absence of serum (CTRL) or in the presence of FBS. Images acquired using an inverted phase-contrast microscope (Axiovert 200) and Axiovision 4.8 software (Zeiss). Scale bar, 100 μm or 50 μm in panels with higher magnification. Periodic acid—Schiff staining performed on HUVECs and Sarc cells (panels on the right) effectively shows that the newly formed tubes are rich in PAS-positive glycogen. ( B ) Representative images of SW1353 cell line and patient-derived ChS cells (ChS-1, ChS-2, and ChS-3), seeded onto GFR-Matrigel and allowed to form vascular-like tubules. Images acquired using an inverted phase-contrast microscope (Axiovert 200) and Axiovision 4.8 software (Zeiss). Scale bar, 500 μm.

    Article Snippet: The SW1353 chondrosarcoma cell line, provided by ATCC, was cultured in Leibovitz’s L-15 Medium (Gibco) with the addition of 10% FBS, 100 μg/mL streptomycin and 100 IU/mL penicillin and maintained at 37 °C, 100% air, according to the manufacturer’s instructions.

    Techniques: In Vitro, Microscopy, Software, Staining, Derivative Assay

    ChS cells do not express VEGFR-2, and VEGF-A does not trigger Sarc cell proliferation. ( A ) Cell lysates (40 µg/samples) from the SW1353 cell line, patient-derived ChS cells, stabilized Sarc cells and HUVECs were resolved on a 10% SDS-PAGE under reducing conditions and transferred onto a nitrocellulose membrane, followed by Western blot with anti-VEGFR-2 and anti-GAPDH as loading control. Chemiluminescent signals were detected and images captured using the eBright 1500 imaging system. ( B , C ) Time-dependent Sarc cell ( B ) and HUVEC ( C ) proliferation monitored using the xCELLigence RTCA technology. Briefly, 1 × 10 4 Sarc cells ( B ) or HUVECs ( C ), seeded in E-16-well plates in the absence (None, in red) or in the presence of 50 ng/mL VEGF-A (VEGF, in blue), were allowed to proliferate over 96 h. The histograms (right panels) report the slope analysis (time range 0–96 h). Data represent mean ± SD from quadruplicate experiments. Student’s t -test ns, not significant; *** p < 0.001.

    Journal: Cells

    Article Title: Vasculogenic Mimicry: A Potential Therapeutic Target for Chondrosarcoma Therapy

    doi: 10.3390/cells15050392

    Figure Lengend Snippet: ChS cells do not express VEGFR-2, and VEGF-A does not trigger Sarc cell proliferation. ( A ) Cell lysates (40 µg/samples) from the SW1353 cell line, patient-derived ChS cells, stabilized Sarc cells and HUVECs were resolved on a 10% SDS-PAGE under reducing conditions and transferred onto a nitrocellulose membrane, followed by Western blot with anti-VEGFR-2 and anti-GAPDH as loading control. Chemiluminescent signals were detected and images captured using the eBright 1500 imaging system. ( B , C ) Time-dependent Sarc cell ( B ) and HUVEC ( C ) proliferation monitored using the xCELLigence RTCA technology. Briefly, 1 × 10 4 Sarc cells ( B ) or HUVECs ( C ), seeded in E-16-well plates in the absence (None, in red) or in the presence of 50 ng/mL VEGF-A (VEGF, in blue), were allowed to proliferate over 96 h. The histograms (right panels) report the slope analysis (time range 0–96 h). Data represent mean ± SD from quadruplicate experiments. Student’s t -test ns, not significant; *** p < 0.001.

    Article Snippet: The SW1353 chondrosarcoma cell line, provided by ATCC, was cultured in Leibovitz’s L-15 Medium (Gibco) with the addition of 10% FBS, 100 μg/mL streptomycin and 100 IU/mL penicillin and maintained at 37 °C, 100% air, according to the manufacturer’s instructions.

    Techniques: Derivative Assay, SDS Page, Membrane, Western Blot, Control, Imaging

    Flow cytometry analysis of vasculogenic-related markers on ChS cell surfaces. The expression levels of vasculogenic markers on SW1353, patient-derived ChS-1, ChS-2, ChS-3, stabilized Sarc ChS cells and HUVECs (with the last cell type used for comparison) were determined by flow cytometry using anti-CD34_PE-Cy7 (sky blue), anti-Podoplanin_ABflo 488 (red), anti-CD31_ABflo 647 (blue), anti-VE Cadherin_ABflo 488 (magenta), anti-EphA2-Alexa 488 (yellow) and anti-uPAR_APC (green). Unstained cells or omission of the primary antibody (when using unconjugated anti-EphA2) were used as negative controls (black lines).

    Journal: Cells

    Article Title: Vasculogenic Mimicry: A Potential Therapeutic Target for Chondrosarcoma Therapy

    doi: 10.3390/cells15050392

    Figure Lengend Snippet: Flow cytometry analysis of vasculogenic-related markers on ChS cell surfaces. The expression levels of vasculogenic markers on SW1353, patient-derived ChS-1, ChS-2, ChS-3, stabilized Sarc ChS cells and HUVECs (with the last cell type used for comparison) were determined by flow cytometry using anti-CD34_PE-Cy7 (sky blue), anti-Podoplanin_ABflo 488 (red), anti-CD31_ABflo 647 (blue), anti-VE Cadherin_ABflo 488 (magenta), anti-EphA2-Alexa 488 (yellow) and anti-uPAR_APC (green). Unstained cells or omission of the primary antibody (when using unconjugated anti-EphA2) were used as negative controls (black lines).

    Article Snippet: The SW1353 chondrosarcoma cell line, provided by ATCC, was cultured in Leibovitz’s L-15 Medium (Gibco) with the addition of 10% FBS, 100 μg/mL streptomycin and 100 IU/mL penicillin and maintained at 37 °C, 100% air, according to the manufacturer’s instructions.

    Techniques: Flow Cytometry, Expressing, Derivative Assay, Comparison

    The inhibitor peptide RI-3 impairs the VM ability of ChS cells. ( A – C ) Representative images of VM by SW1353 ( A ) or patient-derived ChS-1 cells ( C ) suspended in serum free medium (Ctrl-) or 4% FBS medium and seeded onto GFR-Matrigel, in the absence of treatment (None) or in the presence of 10 nM RI-3 peptide or 500 μg/mL Bevacizumab (Beva), and allowed to form vascular-like tubules for 24 h. ( B – D ) The histograms report the quantitative analysis of VM formation performed by the Angiogenesis Analyzer tool of ImageJ software. Data represent mean ± SD from quadruplicate experiments performed at least three times. Student’s t -test ns, not significant, ** p < 0.01, *** p < 0.001.

    Journal: Cells

    Article Title: Vasculogenic Mimicry: A Potential Therapeutic Target for Chondrosarcoma Therapy

    doi: 10.3390/cells15050392

    Figure Lengend Snippet: The inhibitor peptide RI-3 impairs the VM ability of ChS cells. ( A – C ) Representative images of VM by SW1353 ( A ) or patient-derived ChS-1 cells ( C ) suspended in serum free medium (Ctrl-) or 4% FBS medium and seeded onto GFR-Matrigel, in the absence of treatment (None) or in the presence of 10 nM RI-3 peptide or 500 μg/mL Bevacizumab (Beva), and allowed to form vascular-like tubules for 24 h. ( B – D ) The histograms report the quantitative analysis of VM formation performed by the Angiogenesis Analyzer tool of ImageJ software. Data represent mean ± SD from quadruplicate experiments performed at least three times. Student’s t -test ns, not significant, ** p < 0.01, *** p < 0.001.

    Article Snippet: The SW1353 chondrosarcoma cell line, provided by ATCC, was cultured in Leibovitz’s L-15 Medium (Gibco) with the addition of 10% FBS, 100 μg/mL streptomycin and 100 IU/mL penicillin and maintained at 37 °C, 100% air, according to the manufacturer’s instructions.

    Techniques: Derivative Assay, Software