Journal: Molecular Systems Biology

Article Title: Temporal switching and cell-to-cell variability in Ca 2+ release activity in mammalian cells

doi: 10.1038/msb.2009.6

Figure Lengend Snippet: Immunofluorescence detection of RyRs or SERCAs in HEK293 cells. ( A – D ) Upper panels: confocal images showing immunofluorescence localisations of RyRs and SERCAs. Scale bars=20 μm. Lower panels: intercellular histograms of fluorescence intensity in cytoplasmic region. (A) RyR1 was detected using an anti-RyR1 antibody. (B) RyR1 and RyR2 were detected. (C) RyR3 was detected. (D) All SERCA subtypes were detected. The total numbers of cells analysed in the bottom panels are as follows: 214 in (A), 190 in (B), 241 in (C) and 220 in (D).

Article Snippet: The antibodies against RyR1/2 (34C), RyR3 and SERCA (H-300) were purchased from Sigma Aldrich (St Louis, MO), Chemicon (Pittsburgh, PA) and Santa Cruz Biotechnology (Santa Cruz, CA), respectively.

Techniques: Immunofluorescence, Fluorescence

Journal: Molecular Systems Biology

Article Title: Temporal switching and cell-to-cell variability in Ca 2+ release activity in mammalian cells

doi: 10.1038/msb.2009.6

Figure Lengend Snippet: Mathematical model of intracellular Ca 2+ dynamics. ( A , B ) Response of model when RyR activity or SERCA activity changes. (A) RyR activity was increased in a stepwise manner (0.5 (magenta), 2.5 (blue), 4.5 (green), 6.5 (orange) and 8.5 (red) in s −1 ) at a constant SERCA activity (3 μM s −1 ). (B) SERCA activity was decreased in a stepwise manner (5 (magenta), 4 (blue), 3 (green), 2 (orange) and 1 (red) in μM s −1 ) at a constant RyR activity (4.5 s −1 ). ( C ) The balance between the numbers or activities of RyRs and SERCAs determines Ca 2+ response via RyRs with threshold characteristics.

Article Snippet: The antibodies against RyR1/2 (34C), RyR3 and SERCA (H-300) were purchased from Sigma Aldrich (St Louis, MO), Chemicon (Pittsburgh, PA) and Santa Cruz Biotechnology (Santa Cruz, CA), respectively.

Techniques: Activity Assay

Journal: Communications Biology

Article Title: Proteogenomic landscape and clinical characterization of GH-producing pituitary adenomas/somatotroph pituitary neuroendocrine tumors

doi: 10.1038/s42003-022-04272-1

Figure Lengend Snippet: Correlations between ( a ) somatostatin receptor 2 (SSTR2) protein expression, ( b ) SSTR5 protein expression, ( c ) the SSTR2/5 ratio, ( d ) sarcoplasmic/endoplasmic reticulum calcium ATPase 2 (ATP2A2) protein expression, ( e ) ATP2A1 protein expression, and ( f ) AT-rich interaction domain 5B (ARID5B) protein expression and growth hormone (GH) change ratio by octreotide loading test (%). Data were analyzed by Pearson’s correlation analysis. g ATP2A2, ( h ) ATP2A1, and ( i ) ARID5B protein expression levels in NFPA, GNAS -WT GHoma, and GNAS -mutant (MT) GHoma. All protein expression values were log (base 10) transformed. NFPA: nonfunctional pituitary adenoma/non-functioning pituitary neuroendocrine tumor (PitNETs), GHoma: GH-producing pituitary adenoma/somatotroph Pituitary neuroendocrine tumor (PitNETs). * p < 0.05, ** p < 0.01, ***p < 0.001, ****p < 0.0001 by 1-way ANOVA. a – f n = 34 in GHoma with GNAS WT, n = 26 in GHoma with GNAS MT. g – i n = 45 in NFPA, n = 21 in GHoma with GNAS WT, n = 19 in GHoma with GNAS MT.

Article Snippet: Surgically removed adenoma tissues were evaluated by pathological and immunohistochemical examinations using the following antibodies: GH (Dako, Carpinteria, CA, USA; A0570, 1:10), cytokeratin (CK; CAM 5.2; BD Biosciences, San Jose, CA, USA; 345779, 1:10), SSTR2a (Abcam, Cambridge, UK, ab134152, 1:1000), SIGMAR1 (Sigma-Aldrich, Tokyo, Japan, HPA018002, 1:100), ATP2A2 (Sigma-Aldrich, Tokyo, Japan, HPA062605, 1:300), ARID5B (Sigma-Aldrich, Tokyo, Japan, HPA015037, 1:100), WWC3 (Sigma-Aldrich, Tokyo, Japan, HPA039814, 1:500), and SERINC1 (Sigma-Aldrich, Tokyo, Japan, HPA035738, 1:50).

Techniques: Expressing, Mutagenesis, Transformation Assay

Journal: Communications Biology

Article Title: Proteogenomic landscape and clinical characterization of GH-producing pituitary adenomas/somatotroph pituitary neuroendocrine tumors

doi: 10.1038/s42003-022-04272-1

Figure Lengend Snippet: Sections were stained for ( a ) sigma nonopioid intracellular receptor-1 (SIGMAR1), ( b ) sarcoplasmic/endoplasmic reticulum calcium ATPase 2 (ATP2A2), ( c ) AT-rich interaction domain 5B (ARID5B), ( d ) WWC family member 3 (WWC3), and ( e ) serine incorporator 1 (SERINC1). IHC scoring of ( f ) SIGMAR1, ( g ) ATP2A2, ( h ) ARID5B, ( i ) WWC3 and ( j ) SERINC1 was performed by modifying McCarty’s H-score. NFPA: nonfunctional pituitary adenoma/non-functioning pituitary neuroendocrine tumor (PitNETs), GHoma: GH-producing pituitary adenoma/somatotroph Pituitary neuroendocrine tumor (PitNETs). ** p < 0.01, ***p < 0.001 by 1-way ANOVA. n = 8 in NFPA, n = 11 in GHoma with GNAS WT, n = 12 in GHoma with GNAS MT.

Article Snippet: Surgically removed adenoma tissues were evaluated by pathological and immunohistochemical examinations using the following antibodies: GH (Dako, Carpinteria, CA, USA; A0570, 1:10), cytokeratin (CK; CAM 5.2; BD Biosciences, San Jose, CA, USA; 345779, 1:10), SSTR2a (Abcam, Cambridge, UK, ab134152, 1:1000), SIGMAR1 (Sigma-Aldrich, Tokyo, Japan, HPA018002, 1:100), ATP2A2 (Sigma-Aldrich, Tokyo, Japan, HPA062605, 1:300), ARID5B (Sigma-Aldrich, Tokyo, Japan, HPA015037, 1:100), WWC3 (Sigma-Aldrich, Tokyo, Japan, HPA039814, 1:500), and SERINC1 (Sigma-Aldrich, Tokyo, Japan, HPA035738, 1:50).

Techniques: Staining