Journal: Aging Cell

Article Title: Astrocyte Senescence Impairs Synaptogenesis due to Thrombospondin‐1 Loss

doi: 10.1111/acel.70382

Figure Lengend Snippet: 6‐month‐old ACSA‐2+ SAMP8 primary cultured astrocytes and differentiated astrocytes derived from SAMP8 neural stem cells display loss in TSP‐1 levels. (A) RNA‐seq transcriptional profile of Thbs1 gene in astrocytes, neurons, endothelial cells, oligodendrocytes and microglia in P7 (postnatal day) mice. (B) RNA‐seq transcriptional profile of Thbs1 gene at different stages of development and aging in astrocytes from mouse hippocampus. (C) Representative images of GFAP (green) and TSP‐1 (red) immunostaining in ACSA‐2+ primary astrocytes cultures of the SAMP8 and SAMR1 strains (14 DIV) and in Diff‐Ast SAMP8 and SAMR1 strains (11 DIV). (D, E) Quantification of TSP‐1 protein shows lower levels in SAMP8 ACSA‐2+ astrocytes cultures ( p = 0.021) and in Diff‐Ast SAMP8 ( p = 0.011) than in the control SAMR1 strain. (F) RT‐qPCR of Thbs1 ( p = 0.001) demonstrates lower gene expression levels in Diff‐Ast SAMP8 compared to the control line SAMR1. Three independent experiments per cell type were analyzed ( n = 3). Data are presented as mean ± SEM. Unpaired t ‐test was performed in (D, E). One‐sample t ‐test was done in (F). * p < 0.05, ** p < 0.01, *** p < 0.001. Scale bar: 50 μm. Data in (A) and (B) were obtained from databases produced by the Barres lab (Zhang et al. and Clarke et al. available at https://brainrnaseq.org/ ).

Article Snippet: At 11 DIV, neuron medium was totally replaced by ACM from Ast‐Diff or half replaced by ACSA‐2 + ACM during 3 h. Gabapentin (GBP, MedChemExpress, HY‐A0057) and TSP‐1 (MedChemExpress, HY‐P701325) dosages were based on Cheng et al. ( ) procedures.

Techniques: Cell Culture, Derivative Assay, RNA Sequencing, Immunostaining, Control, Quantitative RT-PCR, Gene Expression, Produced

Journal: Aging Cell

Article Title: Astrocyte Senescence Impairs Synaptogenesis due to Thrombospondin‐1 Loss

doi: 10.1111/acel.70382

Figure Lengend Snippet: The hippocampus of 10‐month‐old SAMP8 mice shows deficiencies in TSP‐1 levels. (A) RT‐qPCR of Thbs1 ( p = 0.022) in SAMR1 ( n = 7 independent animals) and SAMP8 ( n = 8 independent animals). (B) TSP‐1 quantification by ELISA test in 10‐m SAMR1 ( n = 4 independent animals) and SAMP8 ( n = 3 independent animals) mice normalized to total protein levels. (C, D) Immunohistochemical analyses and TSP‐1 intensity quantification in 10‐m SAMR1 and SAMP8 hippocampus are shown. Representative astrocytes from the SLM layer in the hippocampus using GFAP (green) as an astroglial marker, and TSP‐1 (red) are depicted magnified. TSP‐1 signal intensity quantification was performed in each layer of the hippocampus and normalized to the quantification area (μm2). Three independent animals of each strain were analyzed for immunohistochemical analysis ( n = 3). Data are presented as mean ± SEM. Unpaired t ‐test was performed. * p < 0.05 and ** p < 0.01. Scale bar: 100 μm; 10 μm (representative astrocyte).

Article Snippet: At 11 DIV, neuron medium was totally replaced by ACM from Ast‐Diff or half replaced by ACSA‐2 + ACM during 3 h. Gabapentin (GBP, MedChemExpress, HY‐A0057) and TSP‐1 (MedChemExpress, HY‐P701325) dosages were based on Cheng et al. ( ) procedures.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Marker

Journal: Aging Cell

Article Title: Astrocyte Senescence Impairs Synaptogenesis due to Thrombospondin‐1 Loss

doi: 10.1111/acel.70382

Figure Lengend Snippet: The competitive TSP‐1 receptor antagonist gabapentin (GBP) blocks the synaptogenic effect of SAMR1 ACMs. (A and C) Immunostaining of MAP2 (gray), VGLUT1 (red) and PSD95 (green), and quantification of excitatory pre‐ (VGLUT1) and postsynaptic (PSD95) vesicles colocalization in hippocampal neurons treated with ACMs from Diff‐Ast SAMR1 and SAMP8 with or without GBP 32 μM. (B and D) Immunostaining of MAP2 (gray), VGLUT1 (red) and PSD95 (green), and quantification of excitatory pre‐ (VGLUT1) and postsynaptic (PSD95) colocalization vesicles in hippocampal neurons treated with ACMs from ACSA‐2 + SAMR1 and SAMP8 of 6‐m mice with or without GBP 32 μM. Three independent experiments were analyzed per cell type and experimental condition. Data are presented as mean ± SEM. One‐way ANOVA Tukey's multiple comparisons test was performed. * p < 0.05, ** p < 0.01, and *** p < 0.001. Scale bar: 50 μm.

Article Snippet: At 11 DIV, neuron medium was totally replaced by ACM from Ast‐Diff or half replaced by ACSA‐2 + ACM during 3 h. Gabapentin (GBP, MedChemExpress, HY‐A0057) and TSP‐1 (MedChemExpress, HY‐P701325) dosages were based on Cheng et al. ( ) procedures.

Techniques: Immunostaining

Journal: Aging Cell

Article Title: Astrocyte Senescence Impairs Synaptogenesis due to Thrombospondin‐1 Loss

doi: 10.1111/acel.70382

Figure Lengend Snippet: TSP‐1 rescues the synaptogenic function of Diff‐Ast SAMP8 ACM. (A, B) Immunostaining of MAP2 (gray), VGLUT1 (red) and PSD95 (green), and quantification of excitatory pre‐ (VGLUT1) and postsynaptic (PSD95) vesicles colocalization in hippocampal neurons treated with ACM from Diff‐Ast SAMP8 with or without TSP‐1 (250 ng/mL) supplementation. (C, D) Immunostaining of MAP2 (gray), VGLUT1 (red) and PSD95 (green), and quantification of excitatory pre‐ (VGLUT1) and postsynaptic (PSD95) vesicles colocalization in hippocampal neurons treated with ACM from transfected Diff‐Ast SAMP8 overexpressing mThbs1 and GFP, or pcDNA3 and GFP as a control. Three independent experiments were analyzed per cell type and experimental condition. Data are presented as mean ± SEM. One‐way ANOVA Tukey's multiple comparisons test was performed. * p < 0.05 and ** p < 0.01. Scale bar: 50 μm.

Article Snippet: At 11 DIV, neuron medium was totally replaced by ACM from Ast‐Diff or half replaced by ACSA‐2 + ACM during 3 h. Gabapentin (GBP, MedChemExpress, HY‐A0057) and TSP‐1 (MedChemExpress, HY‐P701325) dosages were based on Cheng et al. ( ) procedures.

Techniques: Immunostaining, Transfection, Control

Journal: iScience

Article Title: Blockade of TSP-1/CD47 signal axis promotes donor hematopoietic engraftment by improving SEC/MK niche function

doi: 10.1016/j.isci.2025.111952

Figure Lengend Snippet: Improved donor engraftment frequency and peripheral blood count in Thbs1 −/− recipients (A–E) Thbs1 −/− recipients exhibited improvements similar to Cd47 −/− recipients. In this study (A-C), 1.5 × 10 3 sorted B6-GFP LSK (Lin – Sca-1 + c-kit + ) cells were intravenously injected into 9 Gy irradiated WT and Thbs1 −/− mice, along with 2 × 10 5 decoy BM cells from corresponding recipient congenic mice (WT, n = 4; Thbs1 −/− , n = 7). (A) Frequencies (%, left) and numbers (right) of GFP + donor WBCs in peripheral blood at specific time points (4, 10, 15, 19 weeks) post-transplantation. (B) Percentages of GFP + donor CD11b + myeloid cells (left panel), CD19 + B cells (middle panel), and CD3 + T cells (right panel) at the aforementioned time points post-transplantation. (C) Levels (%) of GFP + donor BMCs at 20 weeks post-transplantation. (D and E) WT, Cd47 −/− , and Thbs1 −/− mice were lethally irradiated (9 Gy TBI) 6 h before transplantation, followed by intravenous injection of 3 × 10 3 LSKs from B6-GFP, with 2 × 10 5 decoy BM cells. Blood cells were collected at 3, 7, 12, 17, and 21 weeks post-transplantation for reconstitution analysis. (D) Frequencies (%; left) and numbers (right) of GFP + donor WBCs in peripheral blood at different time points (3, 7, 12, 17, 21 weeks) post-transplantation. (E) Levels (%; left) and numbers of GFP + donor BMCs (right; 1 femur +1 tibia) at 24 weeks post transplantation ( n = 6 for WT, n = 6 for Cd47 −/− , and n = 5 for Thbs1 −/− recipients). Data presented as mean ± SEM. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001, determined by two-way ANOVA with Šídák’s multiple comparisons test (A and B), Unpaired t test (C), or two-way ANOVA with Tukey’s multiple comparison test (D), or one-way ANOVA with Tukey’s multiple comparison test (E).

Article Snippet: Mouse: B6.129S2- Thbs1 tm1Hyn /J , The Jackson Laboratory , Jax:006141.

Techniques: Injection, Irradiation, Transplantation Assay, Comparison

Journal: iScience

Article Title: Blockade of TSP-1/CD47 signal axis promotes donor hematopoietic engraftment by improving SEC/MK niche function

doi: 10.1016/j.isci.2025.111952

Figure Lengend Snippet: Enhanced donor engraftment in SEC/MK niches in Cd47 −/− and Thbs1 −/− mice (A–D) Hoechst staining (blue signal) indicates nuclei, donor HSPCs were presented in the green channel (GFP + ), megakaryocytes (MK) were labeled with CD41-biotin-Strep-PE and CD31/CD144-AF647 (red channel) highlights vascular morphology. (A) The left column presents whole-mount images of donor engraftment in the diaphysis region of WT, Cd47 −/− , and Thbs1 −/− recipients at 60 h post-HSCT (1× 10 6 Lin – BM cells; intravenously), scale bar = 200 μm. The right column displays magnified areas from the left, focusing on the distribution of donor cells (green) around SEC/MK niches, scale bar = 50 μm. (B) Statistical analysis of implanted donor cells; each symbol represents one montage projection. (C) Histogram analysis of distances between donor cells (GFP + ) and MK, computed using Imaris. (D) Statistical analysis of donor cells (Lin – ) in direct contact (0 μm) with megakaryocytes within the BM. Each symbol representing one montage projection, data was presented as mean ± S.D., ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001, determined by one-way ANOVA with Tukey’s multiple comparison test (B and D). Unpaired t test was used (C).

Article Snippet: Mouse: B6.129S2- Thbs1 tm1Hyn /J , The Jackson Laboratory , Jax:006141.

Techniques: Staining, Labeling, Comparison

Journal: iScience

Article Title: Blockade of TSP-1/CD47 signal axis promotes donor hematopoietic engraftment by improving SEC/MK niche function

doi: 10.1016/j.isci.2025.111952

Figure Lengend Snippet: CXCL12 expression pattern at 24 h post 9 Gy TBI in three genetic mice (A) Representative immunofluorescent images of CXCL12 distribution (green channel) at sinusoidal/megakaryocyte niches in cortical region or mid-plane of WT (top panel), Cd47 −/− (middle panel), and Thbs1 −/− (bottom panel) mice at 24 h post-TBI. White arrows highlight megakaryocytes, and white arrowheads indicate thrombi in the sinusoidal space. Scale bar = 50 μm. For corresponding whole mount immunofluorescent images of the bone marrow sections for all three groups, please refer to Figure S5 . (B) Average CXCL12 expression levels were quantified using ImageJ (left), with each symbol representing a single Z montage image. CXCL12 expression in megakaryocytes (middle), or in thrombi (right) was quantified using Imaris, with data collected from 10 to 12 fields at 20 × objective from 3 mice. (C) CXCL12 concentration in BM supernatant in three genetic mice at 24 h post 9 Gy TBI examined by Elisa ( n = 4). Data were presented as mean ± S.D., ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001, determined by one-way ANOVA with Tukey’s multiple comparison test (B and C).

Article Snippet: Mouse: B6.129S2- Thbs1 tm1Hyn /J , The Jackson Laboratory , Jax:006141.

Techniques: Expressing, Concentration Assay, Enzyme-linked Immunosorbent Assay, Comparison

Journal: iScience

Article Title: Blockade of TSP-1/CD47 signal axis promotes donor hematopoietic engraftment by improving SEC/MK niche function

doi: 10.1016/j.isci.2025.111952

Figure Lengend Snippet: Reduced SEC damage and enhanced regeneration in Cd47 −/− and Thbs1 −/− mice during BMT (A–F) Sinusoidal endothelial cells (SECs) exhibited less damage and improved regeneration in Cd47 −/− or Thbs1 −/− mice compared to WT recipients following bone marrow transplantation (BMT). (A) Immunofluorescence for TSP-1 expression in WT, Cd47 −/− , and Thbs1 −/− mice at 24 h after 9 Gy total body irradiation (TBI). The white circle highlights platelets adhering to SECs (WT, Cd47 −/− ) or distributed in thrombi ( Thbs1 −/− ), and the white arrow points to megakaryocytes (MKs). The left two columns display merged images and subsequent TSP-1 expression (green channel), scale bar = 50 μm. The right two columns show magnified areas from the dotted squares; the third column emphasizes platelets (CD41 + , yellow signal) adhering to damaged endothelial cells (CD31 + CD144 + , red signal), and the last column reveals corresponding TSP-1 expression (green channel) in these platelets, scale bar = 10 μm. (B) TSP-1 expression level quantified using ImageJ; each symbol represents one montage projection. (C) Platelets adherence to sinusoidal endothelial cells was computed using Imaris. Each symbol represents one imaging field under a 20 × objective. (D) Fluorescence images showing donor proliferation and vascular morphology 10 days after BMT. Proliferation indicated by Ki-67 (yellow channel), and angiogenesis shown with CD31/CD144 (red channel). Scale bar = 100 μm. (E) Vascular fraction at 24 h post 9 Gy TBI or 10 days post-BMT in WT, Cd47 −/− , or Thbs1 −/− mice, assessed by quantifying CD31 + CD144 + double-positive vascular area relative to total bone marrow area ( n > 10 projections in each group, 3 mice per group). (F) Donor proliferation (Ki-67 bright signals/GFP + area) at 10 days post-1 × 10 6 Lin – BM cell transplantation in the endosteal or central regions of WT, Cd47 −/− , or Thbs1 −/− cohorts. Each symbol represents a single montage projection. Error bars denote SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001, (B and C) were calculated by one-way ANOVA with Tukey’s multiple comparison tests. (E and F) were calculated by two-way ANOVA with Šídák’s multiple comparisons test.

Article Snippet: Mouse: B6.129S2- Thbs1 tm1Hyn /J , The Jackson Laboratory , Jax:006141.

Techniques: Transplantation Assay, Immunofluorescence, Expressing, Irradiation, Imaging, Fluorescence, Comparison

Journal: iScience

Article Title: Blockade of TSP-1/CD47 signal axis promotes donor hematopoietic engraftment by improving SEC/MK niche function

doi: 10.1016/j.isci.2025.111952

Figure Lengend Snippet:

Article Snippet: Mouse: B6.129S2- Thbs1 tm1Hyn /J , The Jackson Laboratory , Jax:006141.

Techniques: Recombinant, Staining, Cell Isolation, TUNEL Assay, In Situ, Derivative Assay, Enzyme-linked Immunosorbent Assay, Bicinchoninic Acid Protein Assay, Software

Journal: International journal of molecular sciences

Article Title: Mechanism of THBS1 Regulation of MDCK Cell Proliferation and Apoptosis Through TGF-β/Smad Signalling.

doi: 10.3390/ijms26010395

Figure Lengend Snippet: Figure 1. Proliferation- and apoptosis-related gene detection in CL23 cells and M60 cells. (a) Proliferation- and apoptosis-related genes were expressed at the mRNA level in CL23 cells and M60 cells. (b) Proliferation- and apopto sis-related genes were expressed at the protein level in CL23 cells and M60 cells. (c) THBS1 and EPHB2 protein level expression in CL23 cells and tu- mourigenic M60 cells. Differential grey value analysis. * indicates statistically significant difference (*** p < 0.001) and no * indicates no difference.

Article Snippet: To further evaluate the mechanism by which THBS1 regulates MDCK cell proliferation and apoptosis through TGF-β/Smad signalling, we observed the cell growth status with different concentration gradients of the TGF-β/Smad signalling pathway activator SRI-011381 (MCE, Shanghai, China, HY-12075) of THBS1-knockdown cells (THBS1-sh122484) and the inhibitor LY2109761 (MCE, Shanghai, China, HY-100347) of THBS1-overexpressing cells (THBS1-OE).

Techniques: Expressing

Journal: International journal of molecular sciences

Article Title: Mechanism of THBS1 Regulation of MDCK Cell Proliferation and Apoptosis Through TGF-β/Smad Signalling.

doi: 10.3390/ijms26010395

Figure Lengend Snippet: Figure 3. Construction and characterisation of MDCK cell lines stably knocking down and overex- pressing THBS1. (a) Bright field and fluorescence expression of cells after puromycin screening of knockdown control cells THBS1-sh-con puromycin; bright field and fluorescence expression of cells after puromycin screening of knockdown cells THBS1-sh-122484 locus; knockdown cells THBS1-sh- 122485 locus puromycin screened for cell bright field and fluorescence expression; knockdown cells THBS1-sh-122486 locus puromycin screened for cell bright field and fluorescence expression; overex- pression of control cells THBS1-OE-con puromycin screened for cell bright field and fluorescence

Article Snippet: To further evaluate the mechanism by which THBS1 regulates MDCK cell proliferation and apoptosis through TGF-β/Smad signalling, we observed the cell growth status with different concentration gradients of the TGF-β/Smad signalling pathway activator SRI-011381 (MCE, Shanghai, China, HY-12075) of THBS1-knockdown cells (THBS1-sh122484) and the inhibitor LY2109761 (MCE, Shanghai, China, HY-100347) of THBS1-overexpressing cells (THBS1-OE).

Techniques: Stable Transfection, Fluorescence, Expressing, Knockdown, Control

Journal: International journal of molecular sciences

Article Title: Mechanism of THBS1 Regulation of MDCK Cell Proliferation and Apoptosis Through TGF-β/Smad Signalling.

doi: 10.3390/ijms26010395

Figure Lengend Snippet: Figure 4. Effects of knockdown of THBS1 on proliferation, apoptosis, migration, and cell cycle of CL23 cells. (a) Growth curves of stably knocked down THBS1 cell lines. (b) Analysis of migratory ability of stably knocked down THBS1 cell lines. (c) Analysis of apoptotic ability of stably knocked down THBS1 cell lines. (d) Analysis of cell cycle of stably knocked down THBS1 cell lines. * denotes statistically significant difference (* p < 0.05; ** p < 0.01; *** p < 0.001) and no * indicates no difference.

Article Snippet: To further evaluate the mechanism by which THBS1 regulates MDCK cell proliferation and apoptosis through TGF-β/Smad signalling, we observed the cell growth status with different concentration gradients of the TGF-β/Smad signalling pathway activator SRI-011381 (MCE, Shanghai, China, HY-12075) of THBS1-knockdown cells (THBS1-sh122484) and the inhibitor LY2109761 (MCE, Shanghai, China, HY-100347) of THBS1-overexpressing cells (THBS1-OE).

Techniques: Knockdown, Migration, Stable Transfection

Journal: International journal of molecular sciences

Article Title: Mechanism of THBS1 Regulation of MDCK Cell Proliferation and Apoptosis Through TGF-β/Smad Signalling.

doi: 10.3390/ijms26010395

Figure Lengend Snippet: Figure 5. Effects of overexpression of THBS1 on proliferation, apoptosis, migration, and cell cycle of M60 cells. (a) Growth curves of stable overexpression of THBS1 cell lines. (b) Analysis of migration ability of stable overexpression of THBS1 cell lines. (c) Analysis of apoptosis ability of stable overexpression of THBS1 cell lines. (d) Analysis of cell cycle of stable overexpression of THBS1 cell lines. * denotes statistically significant difference (** p < 0.01; *** p < 0.001) and no * indicates no difference.

Article Snippet: To further evaluate the mechanism by which THBS1 regulates MDCK cell proliferation and apoptosis through TGF-β/Smad signalling, we observed the cell growth status with different concentration gradients of the TGF-β/Smad signalling pathway activator SRI-011381 (MCE, Shanghai, China, HY-12075) of THBS1-knockdown cells (THBS1-sh122484) and the inhibitor LY2109761 (MCE, Shanghai, China, HY-100347) of THBS1-overexpressing cells (THBS1-OE).

Techniques: Over Expression, Migration

Journal: International journal of molecular sciences

Article Title: Mechanism of THBS1 Regulation of MDCK Cell Proliferation and Apoptosis Through TGF-β/Smad Signalling.

doi: 10.3390/ijms26010395

Figure Lengend Snippet: Figure 6. Effect of knockdown and overexpression of THBS1 on H1N1 influenza virus replication in MDCK cells. (a,b) Differences in expression of NP and NS1 mRNA levels across time of H1N1 influenza virus infection in stably knocked down THBS1 cell lines. (c,d) Differences in expression of NP protein levels across time of H1N1 influenza virus infection in stably knocked down THBS1 cell lines. (e,f) Stable overexpression of THBS1 cell lines infected with H1N1 influenza virus different time period NP and NS1 mRNA level expression differences. (g,h) Stable overexpression of THBS1 cell lines infected with H1N1 influenza virus different time period NP protein level expression differences. * indicates statistically significant difference (* p < 0.05; *** p < 0.001) and no * indicates no difference.

Article Snippet: To further evaluate the mechanism by which THBS1 regulates MDCK cell proliferation and apoptosis through TGF-β/Smad signalling, we observed the cell growth status with different concentration gradients of the TGF-β/Smad signalling pathway activator SRI-011381 (MCE, Shanghai, China, HY-12075) of THBS1-knockdown cells (THBS1-sh122484) and the inhibitor LY2109761 (MCE, Shanghai, China, HY-100347) of THBS1-overexpressing cells (THBS1-OE).

Techniques: Knockdown, Over Expression, Virus, Expressing, Infection, Stable Transfection

Journal: International journal of molecular sciences

Article Title: Mechanism of THBS1 Regulation of MDCK Cell Proliferation and Apoptosis Through TGF-β/Smad Signalling.

doi: 10.3390/ijms26010395

Figure Lengend Snippet: Figure 7. Differential expression of target genes downstream of PI3K/Akt, P53, and TGF-β/Smad signalling pathways, as well as the predicted THBS1-interacting gene, SCARB2, in M60 and CL23 cells. (a,b) Differential expression of target genes downstream of the PI3K/Akt, P53, and TGF-β/Smad signalling pathways, as well as SCARB2 at mRNA level. (c,d) Differential expression of target genes downstream of the TGF-β/Smad signalling pathway and SCARB2 at the protein level. * indicates statistically significant difference (** p < 0.01; *** p < 0.001) and no * indicates no difference.

Article Snippet: To further evaluate the mechanism by which THBS1 regulates MDCK cell proliferation and apoptosis through TGF-β/Smad signalling, we observed the cell growth status with different concentration gradients of the TGF-β/Smad signalling pathway activator SRI-011381 (MCE, Shanghai, China, HY-12075) of THBS1-knockdown cells (THBS1-sh122484) and the inhibitor LY2109761 (MCE, Shanghai, China, HY-100347) of THBS1-overexpressing cells (THBS1-OE).

Techniques: Quantitative Proteomics

Journal: International journal of molecular sciences

Article Title: Mechanism of THBS1 Regulation of MDCK Cell Proliferation and Apoptosis Through TGF-β/Smad Signalling.

doi: 10.3390/ijms26010395

Figure Lengend Snippet: Figure 8. Effects of knockdown and overexpression of THBS1 on the differential expression of target genes downstream of TGF-β/Smad, PI3K/Akt, P53 signalling pathways, and SCARB2 in MDCK cells. (a,b) Effects of knockdown of THBS1 on the differential expression of target genes downstream of PI3K/Akt, P53, TGF-β/Smad signalling, and the SCARB2 mRNA level expression differences in CL23 cells. (c,d) Differential effects of overexpression of THBS1 on the expression of target genes downstream of PI3K/Akt, P53, TGF-β/Smad signalling, and SCARB2 at the mRNA level in M60 cells. (e) Differential effects of knockdown and overexpression of THBS1 on the expression of target genes downstream of the PI3K/Akt, P53, TGF-β/Smad signalling pathways in MDCK cells, and the SCARB2 protein level expression differences. (f,g) Grey value analysis of knockdown and overexpression of THBS1 on the expression of target genes downstream of the PI3K/Akt, P53, and TGF-β/Smad signalling pathways, as well as SCARB2 protein level in MDCK cells. * indicates statistically significant difference (* p < 0.05; ** p < 0.01; *** p < 0.001) and no * indicates no difference.

Article Snippet: To further evaluate the mechanism by which THBS1 regulates MDCK cell proliferation and apoptosis through TGF-β/Smad signalling, we observed the cell growth status with different concentration gradients of the TGF-β/Smad signalling pathway activator SRI-011381 (MCE, Shanghai, China, HY-12075) of THBS1-knockdown cells (THBS1-sh122484) and the inhibitor LY2109761 (MCE, Shanghai, China, HY-100347) of THBS1-overexpressing cells (THBS1-OE).

Techniques: Knockdown, Over Expression, Quantitative Proteomics, Expressing

Journal: International journal of molecular sciences

Article Title: Mechanism of THBS1 Regulation of MDCK Cell Proliferation and Apoptosis Through TGF-β/Smad Signalling.

doi: 10.3390/ijms26010395

Figure Lengend Snippet: Figure 9. Determination of optimal concentration of TGF-β activator SRI-011381 on THBS1-sh-122484 cells and inhibitor LY2109761 on THBS1-OE cells, and its effect on the expression of target genes,

Article Snippet: To further evaluate the mechanism by which THBS1 regulates MDCK cell proliferation and apoptosis through TGF-β/Smad signalling, we observed the cell growth status with different concentration gradients of the TGF-β/Smad signalling pathway activator SRI-011381 (MCE, Shanghai, China, HY-12075) of THBS1-knockdown cells (THBS1-sh122484) and the inhibitor LY2109761 (MCE, Shanghai, China, HY-100347) of THBS1-overexpressing cells (THBS1-OE).

Techniques: Concentration Assay, Expressing

Journal: International journal of molecular sciences

Article Title: Mechanism of THBS1 Regulation of MDCK Cell Proliferation and Apoptosis Through TGF-β/Smad Signalling.

doi: 10.3390/ijms26010395

Figure Lengend Snippet: Figure 10. Effect of SRI-011381 (5 µg/mL) on the expression of PI3K/Akt, target genes downstream of P53 signalling pathway, SCARB2 in knockdown THBS1 cells (THBS1-sh-122484), and LY2109761 (10 µg/mL) in overexpressing THBS1 (THBS1-OE) cells. (a) SRI-011381 (5 µg/mL) intervention in THBS1-sh-122484 cells showed differential expression of PI3K/Akt, target genes downstream of P53 signalling, and SCARB2 at the mRNA level. (b) LY2109761 (10 µg/mL) intervention in THBS1-OE cells showed differential expression of PI3K/Akt, target genes downstream of P53 signalling, and SCARB2 expression differences at the mRNA level. (c) PI3K/Akt, P53 signalling downstream target genes, and SCARB2 expression differences at the protein level after SRI-011381 (5 µg/mL) intervention in THBS1-sh-122484 cells and LY2109761 (10 µg/mL) intervention in THBS1-OE cells. (d) SRI- 011381 (5 µg/mL) intervention in THBS1-sh-122484 cells after PI3K/Akt, P53 signalling downstream target genes, and SCARB2 expression differences at protein level grey value analysis. (e) LY2109761 (10 µg/mL) intervention in THBS1-OE cells after PI3K/Akt, P53 signalling downstream target genes, and SCARB2 expression difference at protein level grey value analysis. * indicates statistically significant difference (* p < 0.05; *** p < 0.001) and no * indicates no difference.

Article Snippet: To further evaluate the mechanism by which THBS1 regulates MDCK cell proliferation and apoptosis through TGF-β/Smad signalling, we observed the cell growth status with different concentration gradients of the TGF-β/Smad signalling pathway activator SRI-011381 (MCE, Shanghai, China, HY-12075) of THBS1-knockdown cells (THBS1-sh122484) and the inhibitor LY2109761 (MCE, Shanghai, China, HY-100347) of THBS1-overexpressing cells (THBS1-OE).

Techniques: Expressing, Knockdown, Quantitative Proteomics

Journal: International journal of molecular sciences

Article Title: Mechanism of THBS1 Regulation of MDCK Cell Proliferation and Apoptosis Through TGF-β/Smad Signalling.

doi: 10.3390/ijms26010395

Figure Lengend Snippet: Figure 11. Effects of SRI-011381 on knockdown THBS1 cells (THBS1-sh-122484) and LY2109761 on proliferation and migration ability of overexpressing THBS1 cells (THBS1-OE). (a) Growth curve of SRI-011381 intervention knockdown THBS1 cells (THBS1-sh-122484). (b,c) SRI-011381 intervention knockdown THBS1 cells (THBS1-sh-122484) migration ability assay. (d) SRI-011381 intervention knockdown THBS1 cells (THBS1-sh-122484) apoptosis ability assay. (e) LY2109761 intervention overexpression THBS1 cells (THBS1-OE) growth curve. (f,g) LY2109761 intervention overexpression THBS1 cells (THBS1-OE) migration ability assay. (h) LY2109761 intervention overexpression THBS1 cells (THBS1-OE) apoptosis ability assay. * indicates statistically significant difference (** p < 0.01; *** p < 0.001) and no * indicates no difference.

Article Snippet: To further evaluate the mechanism by which THBS1 regulates MDCK cell proliferation and apoptosis through TGF-β/Smad signalling, we observed the cell growth status with different concentration gradients of the TGF-β/Smad signalling pathway activator SRI-011381 (MCE, Shanghai, China, HY-12075) of THBS1-knockdown cells (THBS1-sh122484) and the inhibitor LY2109761 (MCE, Shanghai, China, HY-100347) of THBS1-overexpressing cells (THBS1-OE).

Techniques: Knockdown, Migration, Over Expression