Journal: Journal of proteome research

Article Title: Caspase-9 Is a Positive Regulator of Osteoblastic Cell Migration Identified by diaPASEF Proteomics.

doi: 10.1021/acs.jproteome.3c00641

Figure Lengend Snippet: Figure 5. (A) Protein and (B) mRNA levels of ABHD2 and ADAM15 in parental, mock, and Casp9 KO MC3T3-E1 cells; α-tubulin was used as a loading control for immunoblotting. Data represents means ± SD from at least three independent experiments.

Article Snippet: Cells were lysed, and proteins were resolved by SDS-PAGE and immunoblotted as described previously.46 Blots were probed with CASP-9 (#9508, Cell Signaling Technology, USA), cleaved CASP-3 (#9661, Cell Signaling Technology, USA), ABHD2 (14039-1-AP, Proteintech, Germany), ADAM15 (GTX101599, GeneTex, USA), BST-2 (sc-390719, Santa Cruz Biotechnology, USA; 13560-1-AP, Proteintech, Germany; #60066S, Cell Signaling Technology, USA), or α-tubulin (ab7291, Abcam, UK) specific antibodies and horseradish peroxidase-conjugated mouse or rabbit secondary antibodies (Sigma-Aldrich, USA).

Techniques: Control, Western Blot

Journal: Journal of proteome research

Article Title: Caspase-9 Is a Positive Regulator of Osteoblastic Cell Migration Identified by diaPASEF Proteomics.

doi: 10.1021/acs.jproteome.3c00641

Figure Lengend Snippet: Figure 6. Protein levels of ABHD2 and ADAM15 in MC3T3-E1 cells with inhibited CASP-9 or CASP-3/-7; α-tubulin was used as a loading control.

Article Snippet: Cells were lysed, and proteins were resolved by SDS-PAGE and immunoblotted as described previously.46 Blots were probed with CASP-9 (#9508, Cell Signaling Technology, USA), cleaved CASP-3 (#9661, Cell Signaling Technology, USA), ABHD2 (14039-1-AP, Proteintech, Germany), ADAM15 (GTX101599, GeneTex, USA), BST-2 (sc-390719, Santa Cruz Biotechnology, USA; 13560-1-AP, Proteintech, Germany; #60066S, Cell Signaling Technology, USA), or α-tubulin (ab7291, Abcam, UK) specific antibodies and horseradish peroxidase-conjugated mouse or rabbit secondary antibodies (Sigma-Aldrich, USA).

Techniques: Control

Journal: Journal of proteome research

Article Title: Caspase-9 Is a Positive Regulator of Osteoblastic Cell Migration Identified by diaPASEF Proteomics.

doi: 10.1021/acs.jproteome.3c00641

Figure Lengend Snippet: Figure 7. Immunofluorescent staining of ABHD2 (A, C) and cleaved caspase 9 (clCASP9) (B, D) in the developing mouse frontal limb at prenatal day E15. Both proteins were detected in consecutive sections. Positive signal in green (arrows), nuclei counterstained with DAPI (blue); total magnification ×20 (A, B) or ×40 (C, D). hc, hypertrophic cartilage.

Article Snippet: Cells were lysed, and proteins were resolved by SDS-PAGE and immunoblotted as described previously.46 Blots were probed with CASP-9 (#9508, Cell Signaling Technology, USA), cleaved CASP-3 (#9661, Cell Signaling Technology, USA), ABHD2 (14039-1-AP, Proteintech, Germany), ADAM15 (GTX101599, GeneTex, USA), BST-2 (sc-390719, Santa Cruz Biotechnology, USA; 13560-1-AP, Proteintech, Germany; #60066S, Cell Signaling Technology, USA), or α-tubulin (ab7291, Abcam, UK) specific antibodies and horseradish peroxidase-conjugated mouse or rabbit secondary antibodies (Sigma-Aldrich, USA).

Techniques: Staining

Journal: Oncotarget

Article Title: Suppression of ABHD2 , identified through a functional genomics screen, causes anoikis resistance, chemoresistance and poor prognosis in ovarian cancer

doi: 10.18632/oncotarget.9951

Figure Lengend Snippet: a. Seven human ovarian serous adenocarcinoma cell lines and an immortalized human ovarian surface cell line HOSE/E7, all of which do not grow in soft agar, were used. Following transfection of the shRNA library, only OVCA420 cells formed colonies in soft agar. 43 colonies were successfully expanded. shRNAs were amplified by PCR and we reconstructed 69 different shRNA plasmids. Out of the 69 shRNAs in OVCA420 cells, 11 again generated colonies in soft agar. We then measured mRNA expression of these 11 genes using RT-PCR. Of the 11 shRNAs, shRNAs directed against ABHD2, CYB5R3 and ELAC2 suppressed target gene mRNA expression. b. Left: shRNA- ABHD2 transfected OVCA420 cell colony in soft agar. Black bar, 100 μm. Right: normalized ABHD2 / ACTB mRNA expression analyzed by RT-PCR. (n=3, respectively) c. Left: shRNA- ELAC2 transfected OVCA420 cell colony in soft agar. Right: normalized ELAC2 / ACTB mRNA expression. d. Left: shRNA- CYB5R3 transfected OVCA420 cell colony in soft agar. Right: normalized CYB5R3 / ACTB expression.

Article Snippet: Primary antibodies were anti-ABHD2 rabbit polyclonal antibody (1:250, Abgent AP13083c, San Diego, USA), anti-phospho ERK1/2 rabbit monoclonal antibody (1:1,000, Cell Signal Technology, 197G2, Danver, JAPAN), anti-ERK1/2 rabbit monoclonal antibody (1:1000, Cell Signal Technology, 137F5), anti-phospho p38 MAPK rabbit monoclonal antibody (1:1000, Cell Signal Technology, (D13E1)XP) anti-p38MAPK rabbit monoclonal antibody (1:1000, Cell Signal Technology, (D3F9)XP) and anti-GAPDH mouse monoclonal antibody (1:1000, Abcam, ab8245, Cambridge, UK).

Techniques: Transfection, shRNA, Amplification, Generated, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Oncotarget

Article Title: Suppression of ABHD2 , identified through a functional genomics screen, causes anoikis resistance, chemoresistance and poor prognosis in ovarian cancer

doi: 10.18632/oncotarget.9951

Figure Lengend Snippet: mRNA expression was evaluated using log2 normalized values. a. Comparison of ABHD2 mRNA expression between ovarian cancer tissues and serous borderline tumors (SBT) using gene expression microarray datasets GSE9891 and GSE2109. b. Comparison of ABHD2 mRNA expression between serous adenocarcinoma and non-serous adenocarcinoma in microarray dataset GSE2109. c. Copy number alterations for ABHD2 in TCGA samples. Del; deletion, Amp; Amplification. d. Correlation between ABHD2 copy number and mRNA expression in TCGA specimens. e. Representative ABHD2 immunohistochemistry staining for HGSOC (intensity 0, 1 and 2), normal fallopian tube and SBT are shown. Comparison of H-scores among HGSOC, fallopian tube and SBT. The H-score is calculated as 2x the percentage of the most strongly stained area plus 1x the percentage of the most weakly stained area, imparting a total score ranging from 0 to 200.

Article Snippet: Primary antibodies were anti-ABHD2 rabbit polyclonal antibody (1:250, Abgent AP13083c, San Diego, USA), anti-phospho ERK1/2 rabbit monoclonal antibody (1:1,000, Cell Signal Technology, 197G2, Danver, JAPAN), anti-ERK1/2 rabbit monoclonal antibody (1:1000, Cell Signal Technology, 137F5), anti-phospho p38 MAPK rabbit monoclonal antibody (1:1000, Cell Signal Technology, (D13E1)XP) anti-p38MAPK rabbit monoclonal antibody (1:1000, Cell Signal Technology, (D3F9)XP) and anti-GAPDH mouse monoclonal antibody (1:1000, Abcam, ab8245, Cambridge, UK).

Techniques: Expressing, Microarray, Amplification, Immunohistochemistry, Staining

Journal: Oncotarget

Article Title: Suppression of ABHD2 , identified through a functional genomics screen, causes anoikis resistance, chemoresistance and poor prognosis in ovarian cancer

doi: 10.18632/oncotarget.9951

Figure Lengend Snippet: a. Number of viable control, sh1-OVCA420 and sh2-OVCA420 cells following incubation on ultra-low attachment plates (n=6). b. Representative data showing Annexin V/7-ADD staining following incubation on ultralow attachment plates. c. Comparison of the ratio of the Annexin V(−)/7-ADD(−) fraction (viable cells) and Annexin V(+) fraction (apoptotic cells) between control, sh1 and sh2 cells. d. Number of viable OVCA420-control and OVCA420- ABHD2 cells following incubation on ultra-low attachment plates (n=6). Panels a-c: sh1; sh1-OVCA420, sh2; sh2-OVCA420, control; control-OVCA420; panel d: control; OVCA420-control, ABHD2 ; SKOV3- ABHD2 .

Article Snippet: Primary antibodies were anti-ABHD2 rabbit polyclonal antibody (1:250, Abgent AP13083c, San Diego, USA), anti-phospho ERK1/2 rabbit monoclonal antibody (1:1,000, Cell Signal Technology, 197G2, Danver, JAPAN), anti-ERK1/2 rabbit monoclonal antibody (1:1000, Cell Signal Technology, 137F5), anti-phospho p38 MAPK rabbit monoclonal antibody (1:1000, Cell Signal Technology, (D13E1)XP) anti-p38MAPK rabbit monoclonal antibody (1:1000, Cell Signal Technology, (D3F9)XP) and anti-GAPDH mouse monoclonal antibody (1:1000, Abcam, ab8245, Cambridge, UK).

Techniques: Incubation, Staining

Journal: Oncotarget

Article Title: Suppression of ABHD2 , identified through a functional genomics screen, causes anoikis resistance, chemoresistance and poor prognosis in ovarian cancer

doi: 10.18632/oncotarget.9951

Figure Lengend Snippet: a. Number of viable SKOV3-control and SKOV3- ABHD2 cells following incubation on ultra-low attachment plates (n=6). b. Representative data showing Annexin V/7-ADD staining following incubation on ultralow attachment plates. c. Comparison of the ratio of the Annexin V(−)/7-ADD(−) fraction and Annexin V(+) fraction between SKOV3-control and SKOV3- ABHD2 cells. d. Number of viable control- SKOV3, sh1- SKOV3- ABHD2 and sh2-SKOV3- ABHD2 cells following incubation on ultra-low attachment plates (n=6). e. Representative data showing Annexin V/7-ADD staining following incubation on ultralow attachment plates for 48 hours. f. Comparison of the ratio of the Annexin V(−)/7-ADD(−) fraction and Annexin V(+) fraction among control, sh1-SKOV3- ABHD2 and sh2-SKOV3- ABHD2 cells. Panels a-c: control ; SKOV3-control, ABHD2 ; SKOV3- ABHD2 ; panels d-f: control; control-SKOV3- ABHD2, sh1; sh1-SKOV3- ABHD2, sh2 ; sh2-SKOV3- ABHD2 .

Article Snippet: Primary antibodies were anti-ABHD2 rabbit polyclonal antibody (1:250, Abgent AP13083c, San Diego, USA), anti-phospho ERK1/2 rabbit monoclonal antibody (1:1,000, Cell Signal Technology, 197G2, Danver, JAPAN), anti-ERK1/2 rabbit monoclonal antibody (1:1000, Cell Signal Technology, 137F5), anti-phospho p38 MAPK rabbit monoclonal antibody (1:1000, Cell Signal Technology, (D13E1)XP) anti-p38MAPK rabbit monoclonal antibody (1:1000, Cell Signal Technology, (D3F9)XP) and anti-GAPDH mouse monoclonal antibody (1:1000, Abcam, ab8245, Cambridge, UK).

Techniques: Incubation, Staining

Journal: Oncotarget

Article Title: Suppression of ABHD2 , identified through a functional genomics screen, causes anoikis resistance, chemoresistance and poor prognosis in ovarian cancer

doi: 10.18632/oncotarget.9951

Figure Lengend Snippet: All experiments were performed in triplicate. a. Phosphorylated p38 (P-P38) and phosphorylated ERK1/2 (P-ERK1/2) increased following knockdown of ABHD2 (sh1 and sh2) in OVCA420 cells. On the contrary, P-P38 and P-ERK1/2 decreased following overexpression of ABHD2 in SKOV3 and OVCA420 cells. b. Resistance of OVCA420 cells to anoikis on ultra-low attachment dishes was inhibited by GSK1120212, a specific inhibitor of the ERK1/2 pathway. Reduction of P-ERK1/2 following treatment with GSK1120212 was confirmed by Western blotting. DMSO, vehicle control. Cells were treated with differing doses of GSK1120212 as indicated. c. Resistance of OVCA420 cells to anoikis was inhibited following treatment with SB203580, a specific inhibitor of the the p38MAPK pathway. d. Levels of P-P38 and P-ERK1/2 increased following knockdown of ABHD2 (sh1 and sh2) in SKOV3- ABHD2 cells. e. Resistance to anoikis in sh1-SKOV3- ABHD2 and sh2-SKOV3- ABHD2 cells was inhibited following treatment with 100nM GSK1120212 (GSK) and 30μM SB203580 (SB).”

Article Snippet: Primary antibodies were anti-ABHD2 rabbit polyclonal antibody (1:250, Abgent AP13083c, San Diego, USA), anti-phospho ERK1/2 rabbit monoclonal antibody (1:1,000, Cell Signal Technology, 197G2, Danver, JAPAN), anti-ERK1/2 rabbit monoclonal antibody (1:1000, Cell Signal Technology, 137F5), anti-phospho p38 MAPK rabbit monoclonal antibody (1:1000, Cell Signal Technology, (D13E1)XP) anti-p38MAPK rabbit monoclonal antibody (1:1000, Cell Signal Technology, (D3F9)XP) and anti-GAPDH mouse monoclonal antibody (1:1000, Abcam, ab8245, Cambridge, UK).

Techniques: Over Expression, Western Blot

Journal: Oncotarget

Article Title: Suppression of ABHD2 , identified through a functional genomics screen, causes anoikis resistance, chemoresistance and poor prognosis in ovarian cancer

doi: 10.18632/oncotarget.9951

Figure Lengend Snippet: ABHD2 expression levels and clinicopathological factors

Article Snippet: Primary antibodies were anti-ABHD2 rabbit polyclonal antibody (1:250, Abgent AP13083c, San Diego, USA), anti-phospho ERK1/2 rabbit monoclonal antibody (1:1,000, Cell Signal Technology, 197G2, Danver, JAPAN), anti-ERK1/2 rabbit monoclonal antibody (1:1000, Cell Signal Technology, 137F5), anti-phospho p38 MAPK rabbit monoclonal antibody (1:1000, Cell Signal Technology, (D13E1)XP) anti-p38MAPK rabbit monoclonal antibody (1:1000, Cell Signal Technology, (D3F9)XP) and anti-GAPDH mouse monoclonal antibody (1:1000, Abcam, ab8245, Cambridge, UK).

Techniques: Expressing

Journal: Oncotarget

Article Title: Suppression of ABHD2 , identified through a functional genomics screen, causes anoikis resistance, chemoresistance and poor prognosis in ovarian cancer

doi: 10.18632/oncotarget.9951

Figure Lengend Snippet: a. Differences in survival based on ABHD2 immunohistochemical scores (H-score) in HGSOC. b. Differences in survival based on ABHD2 mRNA expression in GSE9891 (n=285, mostly HGSOC) and GSE3149 (n=146, mostly HGSOC) datasets. Samples were divided into high (greater than the median value) and low (less than the median) expression cases. c. Analysis of HGSOC patients (n=36) from KOV-75 based on ABHD2 mRNA expression. d. Analysis of non-HGSOC patients (n=39) from KOV-75 based on ABHD2 mRNA expression. n.s., not significant.

Article Snippet: Primary antibodies were anti-ABHD2 rabbit polyclonal antibody (1:250, Abgent AP13083c, San Diego, USA), anti-phospho ERK1/2 rabbit monoclonal antibody (1:1,000, Cell Signal Technology, 197G2, Danver, JAPAN), anti-ERK1/2 rabbit monoclonal antibody (1:1000, Cell Signal Technology, 137F5), anti-phospho p38 MAPK rabbit monoclonal antibody (1:1000, Cell Signal Technology, (D13E1)XP) anti-p38MAPK rabbit monoclonal antibody (1:1000, Cell Signal Technology, (D3F9)XP) and anti-GAPDH mouse monoclonal antibody (1:1000, Abcam, ab8245, Cambridge, UK).

Techniques: Immunohistochemical staining, Expressing

Journal: Oncotarget

Article Title: Suppression of ABHD2 , identified through a functional genomics screen, causes anoikis resistance, chemoresistance and poor prognosis in ovarian cancer

doi: 10.18632/oncotarget.9951

Figure Lengend Snippet: a. Representative data showing 7-ADD staining following 24 hour incubation with 10 μM cisplatin. b. The ratio of the 7-AAD negative live OVCA420 cells markedly increased following suppression of ABHD2 (sh1 and sh2) as compared to the control after 24 hour incubation with 10 μM cisplatin (n=3). c. Dose-response curves following incubation of OVCA420 cells with the indicated concentrations of cisplatin for 72 hours (n=6). d. Cisplatin IC50 values increased following suppression of ABHD2 in OVCA420 cells. e. Representative data showing 7-ADD staining following 24 hour incubation with 100 μM Carboplatin. f. The ratio of 7-AAD negative live OVCA420 cells markedly increased following suppression of ABHD2 (sh1 and sh2) as compared to the control following a 24 hour incubation with 100 μM carboplatin (n=3).

Article Snippet: Primary antibodies were anti-ABHD2 rabbit polyclonal antibody (1:250, Abgent AP13083c, San Diego, USA), anti-phospho ERK1/2 rabbit monoclonal antibody (1:1,000, Cell Signal Technology, 197G2, Danver, JAPAN), anti-ERK1/2 rabbit monoclonal antibody (1:1000, Cell Signal Technology, 137F5), anti-phospho p38 MAPK rabbit monoclonal antibody (1:1000, Cell Signal Technology, (D13E1)XP) anti-p38MAPK rabbit monoclonal antibody (1:1000, Cell Signal Technology, (D3F9)XP) and anti-GAPDH mouse monoclonal antibody (1:1000, Abcam, ab8245, Cambridge, UK).

Techniques: Staining, Incubation

Journal: Scientific Reports

Article Title: Characterisation and localisation of the endocannabinoid system components in the adult human testis

doi: 10.1038/s41598-019-49177-y

Figure Lengend Snippet: Immunohistochemical staining for the EC-degrading enzymes ABHD2, FAAH and MGLL in the human testis. The images on the left show a general expression pattern and the images on the right show a higher magnification of an area within a stippled line. All pictures show a low magnification image of negative controls in the right upper corner. Note that the ABHD2 (top images) is expressed in the cytoplasm of all stages of germ cell maturation, except early spermatocytes. FAAH reaction (middle row) is strongest in post-meiotic germ cells, including late spermatocytes and spermatids. MGLL protein (bottom images) is abundant in Sertoli cells. L (leptotene) and P (pachytene) indicate early and late spermatocytes, respectively.

Article Snippet: ABHD2: Monoacylglycerol lipase (α/β-hydrolase domain-containing protein 2) , 1:100 , Citrate , Rabbit , Sigma (HPA005999).

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Scientific Reports

Article Title: Characterisation and localisation of the endocannabinoid system components in the adult human testis

doi: 10.1038/s41598-019-49177-y

Figure Lengend Snippet: Antibodies used in the study, listed in alphabetical order.

Article Snippet: ABHD2: Monoacylglycerol lipase (α/β-hydrolase domain-containing protein 2) , 1:100 , Citrate , Rabbit , Sigma (HPA005999).

Techniques:

Journal: Scientific Reports

Article Title: Characterisation and localisation of the endocannabinoid system components in the adult human testis

doi: 10.1038/s41598-019-49177-y

Figure Lengend Snippet: Summary of the results showing predominant expression pattern of the ECS components in specific testicular cell types, grouped into germ cells and somatic cells. Immunohistochemical (IHC) data are shown in upper lines, with the subcellular protein localisation distinguished as nuclear (N) or cellular (C). Transcriptome data from RNA-sequencing (RNA-seq) for available cell types are listed underneath in italics . Note that the RNA-seq data for spermatocytes did not distinguish any specific stages, and that the expression of the receptor RNA isoforms was studied by qPCR.

Article Snippet: ABHD2: Monoacylglycerol lipase (α/β-hydrolase domain-containing protein 2) , 1:100 , Citrate , Rabbit , Sigma (HPA005999).

Techniques: Expressing, Immunohistochemical staining