adam15  (R&D Systems)

Journal: Scientific reports

Article Title: Mechanical forces trigger invasive behavior in synovial fibroblasts through N-cadherin/ADAM15 -dependent modulation of LncRNA H19.

doi: 10.1038/s41598-025-94012-2

Figure Lengend Snippet: Fig. 1. ADAM15-dependent mechano-induced downregulation of lncRNA H19 is affected by inhibition of p21-activated kinases (PAKs). (A, B) Synovial fibroblasts either silenced with an ADAM15 siRNA or a non-silencing negative siRNA were mechanically strained (1 Hz and 15% elongation) for 1–9 h and H19 levels determined by RT-qPCR. (A) Fold changes of H19 in ADAM15-expressing versus non-expressing cells were calculated using the 2−ΔΔCt. Shown is the mean ± SD of 6 different RASFs. (B) time course of GAPDH- normalized Ct values for H19 upon mechanical strain, showing increase of Ct values (i.e. lower H19 amounts) in ADAM15 expressing cells (black dots) compared to ADAM15 non-expressing cells (open circle) in one representative RASF cell line. (C) RT-qPCR for H19 of RASFs stimulated for 1 and 3 h in the presence of either DMEM medium, or inhibitors for JNK (SP600125), Src family kinases (dasatinib), CAMKII (KN- 93), calmodulin (TFP) and PAKs (IPA-3). **p < 0.005, ***p < 0.0005, Student’s t-test, comparing ADAM15- expressing versus non-expressing cells or inhibitor versus DMEM.

Article Snippet: Lysates from 3 pooled 6-wells (200 μg) were incubated with ADAM15 (R&D Systems, #AF935) or N-cadherin (R&D Systems, #AF6426) antibodies (1 μg/mg lysate, respectively) and 25 μl protein G-conjugated agarose beads (Santa Cruz) under constant agitation overnight at 4 °C.

Techniques: Inhibition, Quantitative RT-PCR, Expressing

Journal: Scientific reports

Article Title: Mechanical forces trigger invasive behavior in synovial fibroblasts through N-cadherin/ADAM15 -dependent modulation of LncRNA H19.

doi: 10.1038/s41598-025-94012-2

Figure Lengend Snippet: Fig. 2. ADAM15- and N-cadherin (NCAD) dependent activation of PAK2 upon mechanical strain. (A–D) Immunoblots for pPAK2. (A) of RASFs seeded at different cell densities and mechanically strained for 30 and 60 min. (B) from strained RASFs with PAK2 silencing (si) and treated with a negative control siRNA (neg). (C, D) immunoblots for pPAK2, ADAM15 and NCAD from RASFs treated with ADAM15 and NCAD siRNA; right panels, showing the mean ± SD of densitometric analysis of 6 different RASF cell lines. ***p < 0.0005, Student’s t-test. GAPDH served as a loading control.

Article Snippet: Lysates from 3 pooled 6-wells (200 μg) were incubated with ADAM15 (R&D Systems, #AF935) or N-cadherin (R&D Systems, #AF6426) antibodies (1 μg/mg lysate, respectively) and 25 μl protein G-conjugated agarose beads (Santa Cruz) under constant agitation overnight at 4 °C.

Techniques: Activation Assay, Western Blot, Negative Control, Control

Journal: Scientific reports

Article Title: Mechanical forces trigger invasive behavior in synovial fibroblasts through N-cadherin/ADAM15 -dependent modulation of LncRNA H19.

doi: 10.1038/s41598-025-94012-2

Figure Lengend Snippet: Fig. 3. Interaction of ADAM15 with N-cadherin (NCAD) in synovial fibroblasts. (A) Co- immunoprecipitations (IP) of RASFs with prior silencing of ADAM15 using siRNA (I) and negative control siRNA (N) using NCAD antibodies (upper panels) or ADAM15 antibodies (lower panels) and detection of ADAM15 or NCAD with the respective antibodies. (B) Analogous to (A) Co-IPs using NCAD and ADAM15 antibodies in chondrocyte cell lines transfected with full length ADAM15 plasmid (+) or with ADAM15 lacking the cytoplasmic tail (Δcyto) and an empty vector (-). Mouse IgG served as a control. Input: 10% total lysate. (C) confocal microscopy of RASFs double immunofluorescently stained for NCAD (green) and ADAM15 (red). White inset indicates area of the magnified merged image. (D) In situ proximity ligation assays for ADAM15 and NCAD, showing direct interaction of both molecules in densely seeded cells only (arrows). Cell nuclei were counterstained with DAPI. Objective 40x, scale bar = 10 μm.

Article Snippet: Lysates from 3 pooled 6-wells (200 μg) were incubated with ADAM15 (R&D Systems, #AF935) or N-cadherin (R&D Systems, #AF6426) antibodies (1 μg/mg lysate, respectively) and 25 μl protein G-conjugated agarose beads (Santa Cruz) under constant agitation overnight at 4 °C.

Techniques: Negative Control, Transfection, Plasmid Preparation, Control, Confocal Microscopy, Staining, In Situ, Ligation

Journal: Scientific reports

Article Title: Mechanical forces trigger invasive behavior in synovial fibroblasts through N-cadherin/ADAM15 -dependent modulation of LncRNA H19.

doi: 10.1038/s41598-025-94012-2

Figure Lengend Snippet: Fig. 5. Binding of PAK2 to ADAM15/NCAD complex at cell membrane is crucially dependent on ADAM15. (A) IPs of strained RASFs using ADAM15 or NCAD antibodies and detection of PAK2 and Nck. TL = total lysate. IgG, mouse IgG served as background control. (B) cell surface biotinylation and enrichment of membrane fractions on streptavidin beads of strained cells with ADAM15-silenced or nonsilenced (N). Contr: non biotinylated cell lysates enriched on streptavidin beads served as background control. (C, D) IPs using ADAM15 antibodies in (C) RASFs with silenced and nonsilenced ADAM15 expression (N) and (D) in chondrocyte cell lines transfected with full length ADAM15 (full-A15) or ADAM15 lacking the cytoplasmic domain (Δcyto). (E, F) analogous to (C, D) IPs using N-cadherin antibodies, showing precipitates of PAK2 and Nck in ADAM15-expressing cells only. (E) right, densitometric evaluation of PAK2 from experiments obtained from 4 donors.

Article Snippet: Lysates from 3 pooled 6-wells (200 μg) were incubated with ADAM15 (R&D Systems, #AF935) or N-cadherin (R&D Systems, #AF6426) antibodies (1 μg/mg lysate, respectively) and 25 μl protein G-conjugated agarose beads (Santa Cruz) under constant agitation overnight at 4 °C.

Techniques: Binding Assay, Membrane, Control, Expressing, Transfection

Journal: Scientific reports

Article Title: Mechanical forces trigger invasive behavior in synovial fibroblasts through N-cadherin/ADAM15 -dependent modulation of LncRNA H19.

doi: 10.1038/s41598-025-94012-2

Figure Lengend Snippet: Fig. 6. Cadherin-11, but not Cadherin-2 (NCAD) is a target of mechano-downregulated H19. (A, B) RASFs with ADAM15-nonsilenced (N) or silenced (I) were mechanically strained and (A) RT-qPCR, fold changes of CDH11 or NCAD from 5 different RASFs and (B) immunoblots for CDH11 and NCAD, right, densitometry for CDH11 and NCAD from 4 different RASFs. (C, D) unstimulated RASFs with H19 silenced versus nonsilenced, (C) RT-qPCR, fold changes of CDH11 and NCAD from 5 different RASFs, (D) immunoblots for CDH11 and NCAD, right, densitometry from 4 different RASFs. (E) RT-qPCR for CDH11 and NCAD from mechanically strained (1 h) RASFs incubated with PAK inhibitor IPA-3 or DMEM medium. (F) densitometric evaluation of immunoblots for CDH11 and NCAD from 4 different RASFs (mean ± SD) treated with DMEM or IPA-3 for 48 h. GAPDH served as loading control. **p < 0.005, ***p < 0.0005, Student’s t-test, comparing DMEM versus inhibitor.

Article Snippet: Lysates from 3 pooled 6-wells (200 μg) were incubated with ADAM15 (R&D Systems, #AF935) or N-cadherin (R&D Systems, #AF6426) antibodies (1 μg/mg lysate, respectively) and 25 μl protein G-conjugated agarose beads (Santa Cruz) under constant agitation overnight at 4 °C.

Techniques: Quantitative RT-PCR, Western Blot, Incubation, Control

Journal: Scientific reports

Article Title: Mechanical forces trigger invasive behavior in synovial fibroblasts through N-cadherin/ADAM15 -dependent modulation of LncRNA H19.

doi: 10.1038/s41598-025-94012-2

Figure Lengend Snippet: Fig. 7. ADAM15/H19-dependent downregulation of miR-130a-3p, which binds to 3’ UTR of cadherin-11 and regulates its expression. (A) RNA pull-downs using transfected miR-130a-3p or scrambled mimics, purification on streptavidin beads and detection of binding using qPCR of H19, CDH11 and NCAD, showing Ct values of 5 different RASFs (mean ± SD). (B) RT-qPCR of miR-130a-3p in ADAM15-expressing or silenced RASFs, showing the mean ± SD of fold changes from 5 different RASFs. (C) RT-qPCR for miR-130a-3p in RASFs silenced with two different H19 siRNAs (I and II) vs. negative control siRNA (N). (D, E) RASFs prior transfected with 130a-3p mimics or scramble were strained and (D) RT-qPCR for CDH11 and NCAD and (E) immunoblots for CDH11 and NCAD; untreated = strained without any treatment; right panel, densitometry for CDH11 from 4 different RASFs. GAPDH served as loading control. **p < 0.005, ***p < 0.0005, Student’s t-test.

Article Snippet: Lysates from 3 pooled 6-wells (200 μg) were incubated with ADAM15 (R&D Systems, #AF935) or N-cadherin (R&D Systems, #AF6426) antibodies (1 μg/mg lysate, respectively) and 25 μl protein G-conjugated agarose beads (Santa Cruz) under constant agitation overnight at 4 °C.

Techniques: Expressing, Transfection, Purification, Binding Assay, Quantitative RT-PCR, Negative Control, Western Blot, Control

Journal: Scientific reports

Article Title: Mechanical forces trigger invasive behavior in synovial fibroblasts through N-cadherin/ADAM15 -dependent modulation of LncRNA H19.

doi: 10.1038/s41598-025-94012-2

Figure Lengend Snippet: Fig. 8. CDH11-mediated increased cell invasion can be blocked by CDH11 silencing and transfection with miR-130a-3p mimics. (A, B) RASFs were treated with H19 siRNA or negative nonsilencing siRNA (N) and (A) MTT-based cell viability assays and (B) cells transmigrated through matrigel were stained with DAPI, the whole transwell photographed and all cells counted. (C) upper panel, invaded cells prior transfected with 130a- 3p mimics, CDH11 siRNA or scramble control, shown is the mean ± SD from 5 different RASFs. (C) lower panel, immunoblots for CDH11 from cells transfected and strained as in upper panel. *p < 0.05, **p < 0.005, ***p < 0.0005, Student’s test. (D) diagram of summarized results: mechanical strain results in NCAD/ADAM15- mediated phosphorylation of PAK2, resulting in the downregulation of lncRNA H19 and miR-130a-3p and subsequent upregulation of CDH11, which in turn promotes an aggressive phenotype: increased cell invasion. Inhibition of PAK signaling by PAK inhibitor IPA-3 blocks mechano-induced H19 downregulation and subsequently CDH11 upregulation. In addition, mechano-induced PAK2 phosphorylation is accompanied by recruitment of SH2/SH3 adapter Nck and PAK2 to the NCAD/ADAM15 complex, but binding of Nck/PAK2 is restricted to the cytoplasmic domain of ADAM15. The mechano-induced redistribution of PAK2 to the cell membrane does not occur when Nck is silenced.

Article Snippet: Lysates from 3 pooled 6-wells (200 μg) were incubated with ADAM15 (R&D Systems, #AF935) or N-cadherin (R&D Systems, #AF6426) antibodies (1 μg/mg lysate, respectively) and 25 μl protein G-conjugated agarose beads (Santa Cruz) under constant agitation overnight at 4 °C.

Techniques: Transfection, Staining, Control, Western Blot, Phospho-proteomics, Inhibition, Binding Assay, Membrane

Journal: Scientific reports

Article Title: Mechanical forces trigger invasive behavior in synovial fibroblasts through N-cadherin/ADAM15 -dependent modulation of LncRNA H19.

doi: 10.1038/s41598-025-94012-2

Figure Lengend Snippet: Fig. 1. ADAM15-dependent mechano-induced downregulation of lncRNA H19 is affected by inhibition of p21-activated kinases (PAKs). (A, B) Synovial fibroblasts either silenced with an ADAM15 siRNA or a non-silencing negative siRNA were mechanically strained (1 Hz and 15% elongation) for 1–9 h and H19 levels determined by RT-qPCR. (A) Fold changes of H19 in ADAM15-expressing versus non-expressing cells were calculated using the 2−ΔΔCt. Shown is the mean ± SD of 6 different RASFs. (B) time course of GAPDH- normalized Ct values for H19 upon mechanical strain, showing increase of Ct values (i.e. lower H19 amounts) in ADAM15 expressing cells (black dots) compared to ADAM15 non-expressing cells (open circle) in one representative RASF cell line. (C) RT-qPCR for H19 of RASFs stimulated for 1 and 3 h in the presence of either DMEM medium, or inhibitors for JNK (SP600125), Src family kinases (dasatinib), CAMKII (KN- 93), calmodulin (TFP) and PAKs (IPA-3). **p < 0.005, ***p < 0.0005, Student’s t-test, comparing ADAM15- expressing versus non-expressing cells or inhibitor versus DMEM.

Article Snippet: For detection of ADAM15 and NCAD, cells (1 × 104) were grown on 8-chamberslides for 48 h, fixed with 4% PFA, permeabilized with PBS/0.1% Triton X-100 for 5 min, blocked with 1% horse serum and incubated with mouse ADAM15 (R&D Systems, #MAB935, 1:50) and sheep-anti N-cadherin antibodies.

Techniques: Inhibition, Quantitative RT-PCR, Expressing

Journal: Scientific reports

Article Title: Mechanical forces trigger invasive behavior in synovial fibroblasts through N-cadherin/ADAM15 -dependent modulation of LncRNA H19.

doi: 10.1038/s41598-025-94012-2

Figure Lengend Snippet: Fig. 2. ADAM15- and N-cadherin (NCAD) dependent activation of PAK2 upon mechanical strain. (A–D) Immunoblots for pPAK2. (A) of RASFs seeded at different cell densities and mechanically strained for 30 and 60 min. (B) from strained RASFs with PAK2 silencing (si) and treated with a negative control siRNA (neg). (C, D) immunoblots for pPAK2, ADAM15 and NCAD from RASFs treated with ADAM15 and NCAD siRNA; right panels, showing the mean ± SD of densitometric analysis of 6 different RASF cell lines. ***p < 0.0005, Student’s t-test. GAPDH served as a loading control.

Article Snippet: For detection of ADAM15 and NCAD, cells (1 × 104) were grown on 8-chamberslides for 48 h, fixed with 4% PFA, permeabilized with PBS/0.1% Triton X-100 for 5 min, blocked with 1% horse serum and incubated with mouse ADAM15 (R&D Systems, #MAB935, 1:50) and sheep-anti N-cadherin antibodies.

Techniques: Activation Assay, Western Blot, Negative Control, Control

Journal: Scientific reports

Article Title: Mechanical forces trigger invasive behavior in synovial fibroblasts through N-cadherin/ADAM15 -dependent modulation of LncRNA H19.

doi: 10.1038/s41598-025-94012-2

Figure Lengend Snippet: Fig. 3. Interaction of ADAM15 with N-cadherin (NCAD) in synovial fibroblasts. (A) Co- immunoprecipitations (IP) of RASFs with prior silencing of ADAM15 using siRNA (I) and negative control siRNA (N) using NCAD antibodies (upper panels) or ADAM15 antibodies (lower panels) and detection of ADAM15 or NCAD with the respective antibodies. (B) Analogous to (A) Co-IPs using NCAD and ADAM15 antibodies in chondrocyte cell lines transfected with full length ADAM15 plasmid (+) or with ADAM15 lacking the cytoplasmic tail (Δcyto) and an empty vector (-). Mouse IgG served as a control. Input: 10% total lysate. (C) confocal microscopy of RASFs double immunofluorescently stained for NCAD (green) and ADAM15 (red). White inset indicates area of the magnified merged image. (D) In situ proximity ligation assays for ADAM15 and NCAD, showing direct interaction of both molecules in densely seeded cells only (arrows). Cell nuclei were counterstained with DAPI. Objective 40x, scale bar = 10 μm.

Article Snippet: For detection of ADAM15 and NCAD, cells (1 × 104) were grown on 8-chamberslides for 48 h, fixed with 4% PFA, permeabilized with PBS/0.1% Triton X-100 for 5 min, blocked with 1% horse serum and incubated with mouse ADAM15 (R&D Systems, #MAB935, 1:50) and sheep-anti N-cadherin antibodies.

Techniques: Negative Control, Transfection, Plasmid Preparation, Control, Confocal Microscopy, Staining, In Situ, Ligation

Journal: Scientific reports

Article Title: Mechanical forces trigger invasive behavior in synovial fibroblasts through N-cadherin/ADAM15 -dependent modulation of LncRNA H19.

doi: 10.1038/s41598-025-94012-2

Figure Lengend Snippet: Fig. 5. Binding of PAK2 to ADAM15/NCAD complex at cell membrane is crucially dependent on ADAM15. (A) IPs of strained RASFs using ADAM15 or NCAD antibodies and detection of PAK2 and Nck. TL = total lysate. IgG, mouse IgG served as background control. (B) cell surface biotinylation and enrichment of membrane fractions on streptavidin beads of strained cells with ADAM15-silenced or nonsilenced (N). Contr: non biotinylated cell lysates enriched on streptavidin beads served as background control. (C, D) IPs using ADAM15 antibodies in (C) RASFs with silenced and nonsilenced ADAM15 expression (N) and (D) in chondrocyte cell lines transfected with full length ADAM15 (full-A15) or ADAM15 lacking the cytoplasmic domain (Δcyto). (E, F) analogous to (C, D) IPs using N-cadherin antibodies, showing precipitates of PAK2 and Nck in ADAM15-expressing cells only. (E) right, densitometric evaluation of PAK2 from experiments obtained from 4 donors.

Article Snippet: For detection of ADAM15 and NCAD, cells (1 × 104) were grown on 8-chamberslides for 48 h, fixed with 4% PFA, permeabilized with PBS/0.1% Triton X-100 for 5 min, blocked with 1% horse serum and incubated with mouse ADAM15 (R&D Systems, #MAB935, 1:50) and sheep-anti N-cadherin antibodies.

Techniques: Binding Assay, Membrane, Control, Expressing, Transfection

Journal: Scientific reports

Article Title: Mechanical forces trigger invasive behavior in synovial fibroblasts through N-cadherin/ADAM15 -dependent modulation of LncRNA H19.

doi: 10.1038/s41598-025-94012-2

Figure Lengend Snippet: Fig. 6. Cadherin-11, but not Cadherin-2 (NCAD) is a target of mechano-downregulated H19. (A, B) RASFs with ADAM15-nonsilenced (N) or silenced (I) were mechanically strained and (A) RT-qPCR, fold changes of CDH11 or NCAD from 5 different RASFs and (B) immunoblots for CDH11 and NCAD, right, densitometry for CDH11 and NCAD from 4 different RASFs. (C, D) unstimulated RASFs with H19 silenced versus nonsilenced, (C) RT-qPCR, fold changes of CDH11 and NCAD from 5 different RASFs, (D) immunoblots for CDH11 and NCAD, right, densitometry from 4 different RASFs. (E) RT-qPCR for CDH11 and NCAD from mechanically strained (1 h) RASFs incubated with PAK inhibitor IPA-3 or DMEM medium. (F) densitometric evaluation of immunoblots for CDH11 and NCAD from 4 different RASFs (mean ± SD) treated with DMEM or IPA-3 for 48 h. GAPDH served as loading control. **p < 0.005, ***p < 0.0005, Student’s t-test, comparing DMEM versus inhibitor.

Article Snippet: For detection of ADAM15 and NCAD, cells (1 × 104) were grown on 8-chamberslides for 48 h, fixed with 4% PFA, permeabilized with PBS/0.1% Triton X-100 for 5 min, blocked with 1% horse serum and incubated with mouse ADAM15 (R&D Systems, #MAB935, 1:50) and sheep-anti N-cadherin antibodies.

Techniques: Quantitative RT-PCR, Western Blot, Incubation, Control

Journal: Scientific reports

Article Title: Mechanical forces trigger invasive behavior in synovial fibroblasts through N-cadherin/ADAM15 -dependent modulation of LncRNA H19.

doi: 10.1038/s41598-025-94012-2

Figure Lengend Snippet: Fig. 7. ADAM15/H19-dependent downregulation of miR-130a-3p, which binds to 3’ UTR of cadherin-11 and regulates its expression. (A) RNA pull-downs using transfected miR-130a-3p or scrambled mimics, purification on streptavidin beads and detection of binding using qPCR of H19, CDH11 and NCAD, showing Ct values of 5 different RASFs (mean ± SD). (B) RT-qPCR of miR-130a-3p in ADAM15-expressing or silenced RASFs, showing the mean ± SD of fold changes from 5 different RASFs. (C) RT-qPCR for miR-130a-3p in RASFs silenced with two different H19 siRNAs (I and II) vs. negative control siRNA (N). (D, E) RASFs prior transfected with 130a-3p mimics or scramble were strained and (D) RT-qPCR for CDH11 and NCAD and (E) immunoblots for CDH11 and NCAD; untreated = strained without any treatment; right panel, densitometry for CDH11 from 4 different RASFs. GAPDH served as loading control. **p < 0.005, ***p < 0.0005, Student’s t-test.

Article Snippet: For detection of ADAM15 and NCAD, cells (1 × 104) were grown on 8-chamberslides for 48 h, fixed with 4% PFA, permeabilized with PBS/0.1% Triton X-100 for 5 min, blocked with 1% horse serum and incubated with mouse ADAM15 (R&D Systems, #MAB935, 1:50) and sheep-anti N-cadherin antibodies.

Techniques: Expressing, Transfection, Purification, Binding Assay, Quantitative RT-PCR, Negative Control, Western Blot, Control

Journal: Scientific reports

Article Title: Mechanical forces trigger invasive behavior in synovial fibroblasts through N-cadherin/ADAM15 -dependent modulation of LncRNA H19.

doi: 10.1038/s41598-025-94012-2

Figure Lengend Snippet: Fig. 8. CDH11-mediated increased cell invasion can be blocked by CDH11 silencing and transfection with miR-130a-3p mimics. (A, B) RASFs were treated with H19 siRNA or negative nonsilencing siRNA (N) and (A) MTT-based cell viability assays and (B) cells transmigrated through matrigel were stained with DAPI, the whole transwell photographed and all cells counted. (C) upper panel, invaded cells prior transfected with 130a- 3p mimics, CDH11 siRNA or scramble control, shown is the mean ± SD from 5 different RASFs. (C) lower panel, immunoblots for CDH11 from cells transfected and strained as in upper panel. *p < 0.05, **p < 0.005, ***p < 0.0005, Student’s test. (D) diagram of summarized results: mechanical strain results in NCAD/ADAM15- mediated phosphorylation of PAK2, resulting in the downregulation of lncRNA H19 and miR-130a-3p and subsequent upregulation of CDH11, which in turn promotes an aggressive phenotype: increased cell invasion. Inhibition of PAK signaling by PAK inhibitor IPA-3 blocks mechano-induced H19 downregulation and subsequently CDH11 upregulation. In addition, mechano-induced PAK2 phosphorylation is accompanied by recruitment of SH2/SH3 adapter Nck and PAK2 to the NCAD/ADAM15 complex, but binding of Nck/PAK2 is restricted to the cytoplasmic domain of ADAM15. The mechano-induced redistribution of PAK2 to the cell membrane does not occur when Nck is silenced.

Article Snippet: For detection of ADAM15 and NCAD, cells (1 × 104) were grown on 8-chamberslides for 48 h, fixed with 4% PFA, permeabilized with PBS/0.1% Triton X-100 for 5 min, blocked with 1% horse serum and incubated with mouse ADAM15 (R&D Systems, #MAB935, 1:50) and sheep-anti N-cadherin antibodies.

Techniques: Transfection, Staining, Control, Western Blot, Phospho-proteomics, Inhibition, Binding Assay, Membrane

Journal: Aging (Albany NY)

Article Title: ADAM15 correlates with prognosis, immune infiltration and apoptosis in hepatocellular carcinoma

doi: 10.18632/aging.203425

Figure Lengend Snippet: Correlation between ADAM15 expression and immune cell surface markers.

Article Snippet: Antibodies used included Anti-ADAM15 mouse monoclonal antibody (1:200; cat. no. sc-365752; Santa Cruz Biotechnology, Inc.), anti-E-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab40772; Abcam), anti-N-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab76011; Abcam), anti-Vimentin rabbit monoclonal antibody (1:1000; cat. no. ab92547; Abcam), anti-Snail rabbit monoclonal antibody (1:1000; cat. no. ab216347; Abcam), anti-ZO-1 rabbit monoclonal antibody (1:1000; cat. no. ab276131; Abcam), anti-Bax rabbit polyclonal antibody (1:1000; cat. no. GB11007; Servicebio), anti-Bcl-2 rabbit polyclonal antibody (1:800; cat. no. AF0060; Beyotime Biotechnology).

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: ADAM15 correlates with prognosis, immune infiltration and apoptosis in hepatocellular carcinoma

doi: 10.18632/aging.203425

Figure Lengend Snippet: Different expression of ADAM15 in HCC tissues and corresponding noncancerous tissues. ( A ) ADAM15 expression in pan-cancer. ( B ) ADAM15 expression was up-regulated in HCC tissues compared with paired noncancerous tissues by the TCGA database. ( C , D ) The level of ADAM15 expression was higher in HCC tissues than that in matched noncancerous tissues by the GEO database (GSE76427 and GSE54236). ( E ) ADAM15 expression in HCC samples and paired noncancerous samples was confirmed by western blot, and higher ADAM15 expression was in HCC tissues compared with matched noncancerous tissues. ( F ) ADAM15 expression in HCC samples and paired noncancerous samples was verified by RT-qPCR, and higher ADAM15 expression was in HCC tissues compared with matched noncancerous tissues. ( G ) ADAM15 expression in HCC samples and paired noncancerous samples was confirmed by IHC, and the value of MOD was higher in HCC tissues compared with matched noncancerous tissues. Magnification, ×200. Scale bar: 50 μm ( H , I , J ) Validation of diagnostic value of ADAM15 overexpression in HCC based on the TCGA and GEO databases.

Article Snippet: Antibodies used included Anti-ADAM15 mouse monoclonal antibody (1:200; cat. no. sc-365752; Santa Cruz Biotechnology, Inc.), anti-E-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab40772; Abcam), anti-N-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab76011; Abcam), anti-Vimentin rabbit monoclonal antibody (1:1000; cat. no. ab92547; Abcam), anti-Snail rabbit monoclonal antibody (1:1000; cat. no. ab216347; Abcam), anti-ZO-1 rabbit monoclonal antibody (1:1000; cat. no. ab276131; Abcam), anti-Bax rabbit polyclonal antibody (1:1000; cat. no. GB11007; Servicebio), anti-Bcl-2 rabbit polyclonal antibody (1:800; cat. no. AF0060; Beyotime Biotechnology).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Biomarker Discovery, Diagnostic Assay, Over Expression

Journal: Aging (Albany NY)

Article Title: ADAM15 correlates with prognosis, immune infiltration and apoptosis in hepatocellular carcinoma

doi: 10.18632/aging.203425

Figure Lengend Snippet: The relationships between ADAM15 expression and clinicopathological parameters in HCC.

Article Snippet: Antibodies used included Anti-ADAM15 mouse monoclonal antibody (1:200; cat. no. sc-365752; Santa Cruz Biotechnology, Inc.), anti-E-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab40772; Abcam), anti-N-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab76011; Abcam), anti-Vimentin rabbit monoclonal antibody (1:1000; cat. no. ab92547; Abcam), anti-Snail rabbit monoclonal antibody (1:1000; cat. no. ab216347; Abcam), anti-ZO-1 rabbit monoclonal antibody (1:1000; cat. no. ab276131; Abcam), anti-Bax rabbit polyclonal antibody (1:1000; cat. no. GB11007; Servicebio), anti-Bcl-2 rabbit polyclonal antibody (1:800; cat. no. AF0060; Beyotime Biotechnology).

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: ADAM15 correlates with prognosis, immune infiltration and apoptosis in hepatocellular carcinoma

doi: 10.18632/aging.203425

Figure Lengend Snippet: Survival analysis and prognostic value of ADAM15 between different groups. ( A ) Kaplan-Meier survival analysis of OS in HCC. ( B ) Kaplan-Meier survival analysis of RFS in HCC. ( C ) Univariate Cox regression analysis of OS in HCC based on TCGA database. ( D ) Multivariate Cox regression analysis of OS in HCC based on TCGA database. ( E ) Univariate Cox regression analysis of RFS in HCC based on TCGA database. ( F ) Multivariate Cox regression analysis of RFS in HCC based on TCGA database.

Article Snippet: Antibodies used included Anti-ADAM15 mouse monoclonal antibody (1:200; cat. no. sc-365752; Santa Cruz Biotechnology, Inc.), anti-E-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab40772; Abcam), anti-N-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab76011; Abcam), anti-Vimentin rabbit monoclonal antibody (1:1000; cat. no. ab92547; Abcam), anti-Snail rabbit monoclonal antibody (1:1000; cat. no. ab216347; Abcam), anti-ZO-1 rabbit monoclonal antibody (1:1000; cat. no. ab276131; Abcam), anti-Bax rabbit polyclonal antibody (1:1000; cat. no. GB11007; Servicebio), anti-Bcl-2 rabbit polyclonal antibody (1:800; cat. no. AF0060; Beyotime Biotechnology).

Techniques:

Journal: Aging (Albany NY)

Article Title: ADAM15 correlates with prognosis, immune infiltration and apoptosis in hepatocellular carcinoma

doi: 10.18632/aging.203425

Figure Lengend Snippet: The prognostic value of ADAM15 was validated by nomograms. ( A ) The nomogram of OS. ( B ) The 1-year survival rate of OS. ( C ) The 3-year survival rate of OS. ( D ) The 5-year survival rate of OS. ( E ) The nomogram of RFS. ( F ) The 1-year survival rate of RFS. ( G ) The 3-year survival rate of RFS. ( H ) The 5-year survival rate of RFS.

Article Snippet: Antibodies used included Anti-ADAM15 mouse monoclonal antibody (1:200; cat. no. sc-365752; Santa Cruz Biotechnology, Inc.), anti-E-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab40772; Abcam), anti-N-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab76011; Abcam), anti-Vimentin rabbit monoclonal antibody (1:1000; cat. no. ab92547; Abcam), anti-Snail rabbit monoclonal antibody (1:1000; cat. no. ab216347; Abcam), anti-ZO-1 rabbit monoclonal antibody (1:1000; cat. no. ab276131; Abcam), anti-Bax rabbit polyclonal antibody (1:1000; cat. no. GB11007; Servicebio), anti-Bcl-2 rabbit polyclonal antibody (1:800; cat. no. AF0060; Beyotime Biotechnology).

Techniques:

Journal: Aging (Albany NY)

Article Title: ADAM15 correlates with prognosis, immune infiltration and apoptosis in hepatocellular carcinoma

doi: 10.18632/aging.203425

Figure Lengend Snippet: ADAM15 expression associated with immune infiltration in HCC. ( A , B ) The relationship between ADAM15 expression and immune infiltration cells confirmed by TIMER and TISIDB databases. ( C ) ADAM15 is highly expressed in the high immune score group based on the TCGA database. ( D ) Kaplan-Meier survival analysis of OS in the low immune score group of HCC based on the TCGA database. ( E ) Kaplan-Meier survival analysis of OS in the high immune score group of HCC based on the TCGA database. ( F ) ADAM15 is highly expressed in high stromal score group based on the TCGA database. ( G ) Kaplan-Meier survival analysis of OS in the low stromal score group of HCC based on the TCGA database. ( H ) Kaplan-Meier survival analysis of OS in the high stromal score group of HCC based on the TCGA database.

Article Snippet: Antibodies used included Anti-ADAM15 mouse monoclonal antibody (1:200; cat. no. sc-365752; Santa Cruz Biotechnology, Inc.), anti-E-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab40772; Abcam), anti-N-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab76011; Abcam), anti-Vimentin rabbit monoclonal antibody (1:1000; cat. no. ab92547; Abcam), anti-Snail rabbit monoclonal antibody (1:1000; cat. no. ab216347; Abcam), anti-ZO-1 rabbit monoclonal antibody (1:1000; cat. no. ab276131; Abcam), anti-Bax rabbit polyclonal antibody (1:1000; cat. no. GB11007; Servicebio), anti-Bcl-2 rabbit polyclonal antibody (1:800; cat. no. AF0060; Beyotime Biotechnology).

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: ADAM15 correlates with prognosis, immune infiltration and apoptosis in hepatocellular carcinoma

doi: 10.18632/aging.203425

Figure Lengend Snippet: ADAM15 expression was correlated with markers of immune cells. ( A ) B cell markers, CD19 and CD38. ( B ) CD4+T cell markers, IL2RA and CTLA4. ( C ) CD8+T cell, CD8A and CD8B. ( D ) Macrophage markers, ITGAM and CD68. ( E ) Neutrophil markers, FCGR3A and FUT4. ( F ) DC markers, ITGAE and THBD.

Article Snippet: Antibodies used included Anti-ADAM15 mouse monoclonal antibody (1:200; cat. no. sc-365752; Santa Cruz Biotechnology, Inc.), anti-E-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab40772; Abcam), anti-N-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab76011; Abcam), anti-Vimentin rabbit monoclonal antibody (1:1000; cat. no. ab92547; Abcam), anti-Snail rabbit monoclonal antibody (1:1000; cat. no. ab216347; Abcam), anti-ZO-1 rabbit monoclonal antibody (1:1000; cat. no. ab276131; Abcam), anti-Bax rabbit polyclonal antibody (1:1000; cat. no. GB11007; Servicebio), anti-Bcl-2 rabbit polyclonal antibody (1:800; cat. no. AF0060; Beyotime Biotechnology).

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: ADAM15 correlates with prognosis, immune infiltration and apoptosis in hepatocellular carcinoma

doi: 10.18632/aging.203425

Figure Lengend Snippet: The association of ADAM15 expression with immune checkpoint genes. ( A ) The differential expression of immune checkpoint genes between the high and low expression group. ( B ) The survival analysis of CD80. ( B ) The survival analysis of LDHA. ( C ) The survival analysis of TNFRSF4. ( D ) The survival analysis of TNFSF4. ( E ) The survival analysis of TNFSF4. ( F ) The survival analysis of YTHDF1.

Article Snippet: Antibodies used included Anti-ADAM15 mouse monoclonal antibody (1:200; cat. no. sc-365752; Santa Cruz Biotechnology, Inc.), anti-E-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab40772; Abcam), anti-N-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab76011; Abcam), anti-Vimentin rabbit monoclonal antibody (1:1000; cat. no. ab92547; Abcam), anti-Snail rabbit monoclonal antibody (1:1000; cat. no. ab216347; Abcam), anti-ZO-1 rabbit monoclonal antibody (1:1000; cat. no. ab276131; Abcam), anti-Bax rabbit polyclonal antibody (1:1000; cat. no. GB11007; Servicebio), anti-Bcl-2 rabbit polyclonal antibody (1:800; cat. no. AF0060; Beyotime Biotechnology).

Techniques: Expressing, Quantitative Proteomics

Journal: Aging (Albany NY)

Article Title: ADAM15 correlates with prognosis, immune infiltration and apoptosis in hepatocellular carcinoma

doi: 10.18632/aging.203425

Figure Lengend Snippet: The level of ADAM15 expression was associated with the proliferation, migration and invasion of HCC cells. ( A ) Comparison of ADAM15 expression in HCC cells and liver cells. ( B , C ) Cell proliferation ability of HepG2 and SMMC-7721 cell in siCon group, siADAM15 group and ADAM15 overexpression group was examined by CCK8 assay. ( D ) Overexpression of ADAM15 was evaluated by western blot in HepG2 cell. ( E ) ADAM15 knockdown was assessed by western blot in SMMC-7721 cell. ( F ) Cell migration ability of HepG2 and SMMC-7721 cell in siCon group, siADAM15 group and ADAM15 overexpression group was examined by a wound-healing method. Magnification, x40. Scale bar: 200 μm ( G ) Cell invasion ability of HepG2 and SMMC-7721 cell in siCon group, siADAM15 group and ADAM15 overexpression group was evaluated by transwell assay. Magnification, x200. Scale bar: 50 μm.

Article Snippet: Antibodies used included Anti-ADAM15 mouse monoclonal antibody (1:200; cat. no. sc-365752; Santa Cruz Biotechnology, Inc.), anti-E-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab40772; Abcam), anti-N-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab76011; Abcam), anti-Vimentin rabbit monoclonal antibody (1:1000; cat. no. ab92547; Abcam), anti-Snail rabbit monoclonal antibody (1:1000; cat. no. ab216347; Abcam), anti-ZO-1 rabbit monoclonal antibody (1:1000; cat. no. ab276131; Abcam), anti-Bax rabbit polyclonal antibody (1:1000; cat. no. GB11007; Servicebio), anti-Bcl-2 rabbit polyclonal antibody (1:800; cat. no. AF0060; Beyotime Biotechnology).

Techniques: Expressing, Migration, Comparison, Over Expression, CCK-8 Assay, Western Blot, Knockdown, Transwell Assay

Journal: Aging (Albany NY)

Article Title: ADAM15 correlates with prognosis, immune infiltration and apoptosis in hepatocellular carcinoma

doi: 10.18632/aging.203425

Figure Lengend Snippet: Impacts of ADAM15 on the expression of apoptosis-related proteins and EMT-related proteins. ( A ) Functional enrichment analyses indicated that overexpression of ADAM15 was associated with apoptosis. ( B ) Functional enrichment analyses indicated that overexpression of ADAM15 was related to focal adhesion. ( C ) Flow cytometry analysis indicated that the siRNA-transfected in SMMC-7721 cells significantly induced apoptosis. ( D ) Flow cytometry analysis indicated that the overexpression of ADAM15 in HepG2 cells significantly suppressed apoptosis. ( E ) Comparison of apoptosis-related protein expression between the control group and siRNA3 group in SMMC-7721 cells. ( F ) Comparison of apoptosis-related protein expression between the control group and overexpression of ADAM15 group in HepG2 cell. ( G ) Comparison of EMT-related protein expression between the control group and siRNA3 group in SMMC-7721 cells. ( H ) Comparison of EMT-related protein expression between the control group and overexpression of ADAM15 group in HepG2 cells.

Article Snippet: Antibodies used included Anti-ADAM15 mouse monoclonal antibody (1:200; cat. no. sc-365752; Santa Cruz Biotechnology, Inc.), anti-E-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab40772; Abcam), anti-N-cadherin rabbit monoclonal antibody (1:1000; cat. no. ab76011; Abcam), anti-Vimentin rabbit monoclonal antibody (1:1000; cat. no. ab92547; Abcam), anti-Snail rabbit monoclonal antibody (1:1000; cat. no. ab216347; Abcam), anti-ZO-1 rabbit monoclonal antibody (1:1000; cat. no. ab276131; Abcam), anti-Bax rabbit polyclonal antibody (1:1000; cat. no. GB11007; Servicebio), anti-Bcl-2 rabbit polyclonal antibody (1:800; cat. no. AF0060; Beyotime Biotechnology).

Techniques: Expressing, Functional Assay, Over Expression, Flow Cytometry, Transfection, Comparison, Control