sw1353 (ATCC)
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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
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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
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
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
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
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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