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Fig. 5 <t>TIMP3</t> negatively regulates glycosaminoglycan-dependent BG ectodomain shedding. A Scatter plot of the differentially expressed genes from transcriptomics of HS-BG and ∆GAG-BG plotted against CS-BG and ∆GAG-BG comparison groups. All protease-related genes are highlighted. B Volcano plots of the differentially expressed genes between ∆GAG-BG vs. Control-vector, S534A (CS-BG) vs. ∆GAG-BG, and S545A (HS-BG) vs. ∆GAG-BG. (TIMP3; ∆GAG vs Cntl, log2FC = 0.524, -log(10)p = 1.643, ∆GAG vs CS-BG, log2FC = 1.054, -log(10)p = 2.493, ∆GAG vs HS-BG, log2FC = 0.364, -log(10)p = 1.1411.). C, E Semi-qRT-PCR of TIMP3 and (D, F) ELISA for sol-BG in indicated cells normalized to the scramble vector cells. Mean ± SEM (n = 3 qRT-PCR), *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001, One-way ANOVA followed by unpaired t-test between scramble vector treated cells to siTIMP3 treated cells. (HEYA8 p = 0.0055 ∆GAG-BG SCRM vs siTIMP3, SKOV-3 p = 0.0007, ∆GAG-BG SCRM compared to siTIMP3), for ELISA in (D, F) n = 5 HEYA8, n = 6 SKOV-3, p = 0.0068, ∆GAG-BG SCRM vs siTIMP3 in HEYA8 and p = < 0.0001, ∆GAG SCRM vs. siTIMP3 in SKOV-3
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Fig. 5 <t>TIMP3</t> negatively regulates glycosaminoglycan-dependent BG ectodomain shedding. A Scatter plot of the differentially expressed genes from transcriptomics of HS-BG and ∆GAG-BG plotted against CS-BG and ∆GAG-BG comparison groups. All protease-related genes are highlighted. B Volcano plots of the differentially expressed genes between ∆GAG-BG vs. Control-vector, S534A (CS-BG) vs. ∆GAG-BG, and S545A (HS-BG) vs. ∆GAG-BG. (TIMP3; ∆GAG vs Cntl, log2FC = 0.524, -log(10)p = 1.643, ∆GAG vs CS-BG, log2FC = 1.054, -log(10)p = 2.493, ∆GAG vs HS-BG, log2FC = 0.364, -log(10)p = 1.1411.). C, E Semi-qRT-PCR of TIMP3 and (D, F) ELISA for sol-BG in indicated cells normalized to the scramble vector cells. Mean ± SEM (n = 3 qRT-PCR), *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001, One-way ANOVA followed by unpaired t-test between scramble vector treated cells to siTIMP3 treated cells. (HEYA8 p = 0.0055 ∆GAG-BG SCRM vs siTIMP3, SKOV-3 p = 0.0007, ∆GAG-BG SCRM compared to siTIMP3), for ELISA in (D, F) n = 5 HEYA8, n = 6 SKOV-3, p = 0.0068, ∆GAG-BG SCRM vs siTIMP3 in HEYA8 and p = < 0.0001, ∆GAG SCRM vs. siTIMP3 in SKOV-3
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Fig. 5 <t>TIMP3</t> negatively regulates glycosaminoglycan-dependent BG ectodomain shedding. A Scatter plot of the differentially expressed genes from transcriptomics of HS-BG and ∆GAG-BG plotted against CS-BG and ∆GAG-BG comparison groups. All protease-related genes are highlighted. B Volcano plots of the differentially expressed genes between ∆GAG-BG vs. Control-vector, S534A (CS-BG) vs. ∆GAG-BG, and S545A (HS-BG) vs. ∆GAG-BG. (TIMP3; ∆GAG vs Cntl, log2FC = 0.524, -log(10)p = 1.643, ∆GAG vs CS-BG, log2FC = 1.054, -log(10)p = 2.493, ∆GAG vs HS-BG, log2FC = 0.364, -log(10)p = 1.1411.). C, E Semi-qRT-PCR of TIMP3 and (D, F) ELISA for sol-BG in indicated cells normalized to the scramble vector cells. Mean ± SEM (n = 3 qRT-PCR), *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001, One-way ANOVA followed by unpaired t-test between scramble vector treated cells to siTIMP3 treated cells. (HEYA8 p = 0.0055 ∆GAG-BG SCRM vs siTIMP3, SKOV-3 p = 0.0007, ∆GAG-BG SCRM compared to siTIMP3), for ELISA in (D, F) n = 5 HEYA8, n = 6 SKOV-3, p = 0.0068, ∆GAG-BG SCRM vs siTIMP3 in HEYA8 and p = < 0.0001, ∆GAG SCRM vs. siTIMP3 in SKOV-3
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Fig. 5 <t>TIMP3</t> negatively regulates glycosaminoglycan-dependent BG ectodomain shedding. A Scatter plot of the differentially expressed genes from transcriptomics of HS-BG and ∆GAG-BG plotted against CS-BG and ∆GAG-BG comparison groups. All protease-related genes are highlighted. B Volcano plots of the differentially expressed genes between ∆GAG-BG vs. Control-vector, S534A (CS-BG) vs. ∆GAG-BG, and S545A (HS-BG) vs. ∆GAG-BG. (TIMP3; ∆GAG vs Cntl, log2FC = 0.524, -log(10)p = 1.643, ∆GAG vs CS-BG, log2FC = 1.054, -log(10)p = 2.493, ∆GAG vs HS-BG, log2FC = 0.364, -log(10)p = 1.1411.). C, E Semi-qRT-PCR of TIMP3 and (D, F) ELISA for sol-BG in indicated cells normalized to the scramble vector cells. Mean ± SEM (n = 3 qRT-PCR), *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001, One-way ANOVA followed by unpaired t-test between scramble vector treated cells to siTIMP3 treated cells. (HEYA8 p = 0.0055 ∆GAG-BG SCRM vs siTIMP3, SKOV-3 p = 0.0007, ∆GAG-BG SCRM compared to siTIMP3), for ELISA in (D, F) n = 5 HEYA8, n = 6 SKOV-3, p = 0.0068, ∆GAG-BG SCRM vs siTIMP3 in HEYA8 and p = < 0.0001, ∆GAG SCRM vs. siTIMP3 in SKOV-3
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Fig. 5 TIMP3 negatively regulates glycosaminoglycan-dependent BG ectodomain shedding. A Scatter plot of the differentially expressed genes from transcriptomics of HS-BG and ∆GAG-BG plotted against CS-BG and ∆GAG-BG comparison groups. All protease-related genes are highlighted. B Volcano plots of the differentially expressed genes between ∆GAG-BG vs. Control-vector, S534A (CS-BG) vs. ∆GAG-BG, and S545A (HS-BG) vs. ∆GAG-BG. (TIMP3; ∆GAG vs Cntl, log2FC = 0.524, -log(10)p = 1.643, ∆GAG vs CS-BG, log2FC = 1.054, -log(10)p = 2.493, ∆GAG vs HS-BG, log2FC = 0.364, -log(10)p = 1.1411.). C, E Semi-qRT-PCR of TIMP3 and (D, F) ELISA for sol-BG in indicated cells normalized to the scramble vector cells. Mean ± SEM (n = 3 qRT-PCR), *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001, One-way ANOVA followed by unpaired t-test between scramble vector treated cells to siTIMP3 treated cells. (HEYA8 p = 0.0055 ∆GAG-BG SCRM vs siTIMP3, SKOV-3 p = 0.0007, ∆GAG-BG SCRM compared to siTIMP3), for ELISA in (D, F) n = 5 HEYA8, n = 6 SKOV-3, p = 0.0068, ∆GAG-BG SCRM vs siTIMP3 in HEYA8 and p = < 0.0001, ∆GAG SCRM vs. siTIMP3 in SKOV-3

Journal: Cell communication and signaling : CCS

Article Title: Glycosaminoglycan modifications of betaglycan regulate ectodomain shedding to fine-tune TGF-β signaling responses in ovarian cancer.

doi: 10.1186/s12964-024-01496-y

Figure Lengend Snippet: Fig. 5 TIMP3 negatively regulates glycosaminoglycan-dependent BG ectodomain shedding. A Scatter plot of the differentially expressed genes from transcriptomics of HS-BG and ∆GAG-BG plotted against CS-BG and ∆GAG-BG comparison groups. All protease-related genes are highlighted. B Volcano plots of the differentially expressed genes between ∆GAG-BG vs. Control-vector, S534A (CS-BG) vs. ∆GAG-BG, and S545A (HS-BG) vs. ∆GAG-BG. (TIMP3; ∆GAG vs Cntl, log2FC = 0.524, -log(10)p = 1.643, ∆GAG vs CS-BG, log2FC = 1.054, -log(10)p = 2.493, ∆GAG vs HS-BG, log2FC = 0.364, -log(10)p = 1.1411.). C, E Semi-qRT-PCR of TIMP3 and (D, F) ELISA for sol-BG in indicated cells normalized to the scramble vector cells. Mean ± SEM (n = 3 qRT-PCR), *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001, One-way ANOVA followed by unpaired t-test between scramble vector treated cells to siTIMP3 treated cells. (HEYA8 p = 0.0055 ∆GAG-BG SCRM vs siTIMP3, SKOV-3 p = 0.0007, ∆GAG-BG SCRM compared to siTIMP3), for ELISA in (D, F) n = 5 HEYA8, n = 6 SKOV-3, p = 0.0068, ∆GAG-BG SCRM vs siTIMP3 in HEYA8 and p = < 0.0001, ∆GAG SCRM vs. siTIMP3 in SKOV-3

Article Snippet: Human TGF-beta RIII (catalog AF-242) was purchased from R&D Biosystems (Minneapolis, MN), PhosphoSMAD2 (Ser465/467)/SMAD3 (Ser423/425) (catalog #8828), Smad2/3 (D7G7) (catalog #8685), Smad2 (D43B4) XP® Rabbit mAb (catalog #5339), TIMP3 (D74B10) Rabbit mAb (catalog #5673) were obtained from Cell Signal Technology (Danvers, MA).

Techniques: Comparison, Control, Plasmid Preparation, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay