Journal: International Journal of Molecular Sciences

Article Title: Mouse Ataxin-2 Expansion Downregulates CamKII and Other Calcium Signaling Factors, Impairing Granule—Purkinje Neuron Synaptic Strength

doi: 10.3390/ijms21186673

Figure Lengend Snippet: List of TaqMan ® Assays utilized in this study.

Article Snippet: Atp2a1 , Mm01275320_m1 , Grm4 , Mm01306128_m1 , Pcp4 , Mm00500973_m1.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Diaphragm Fatigue in SMNΔ7 Mice and Its Molecular Determinants: An Underestimated Issue

doi: 10.3390/ijms241914953

Figure Lengend Snippet: Alterations of contractile functions in SMA diaphragm. ( a ) Mean values of twitch time to peak (TTP), twitch half relaxation time (TTR), twitch specific tension (Po/CSA), and specific tetanic tension determined in ex vivo functional analysis in the diaphragm muscles (WT, n = 21; SMA, n = 17). ( b ) Fatigue index (percentage of the maximal tetanic force) (WT, n = 12; SMA, n = 12). ( c ) Gene (by RT-PCR) expression (WT, n = 5; SMA, n = 3) and protein (by Western blot) expression (WT, n = 8; SMA, n = 7) of sarcoplasmic reticulum Ca 2+ ATPase pumps (SERCA1 and 2). The level of protein target was normalized against the level of the housekeeping tubulin measured in the same blot. Representative Western blots are shown. Bars represent means± SD. Individual data are represented as scatter plots. * p ≤ 0.05 ** p ≤ 0.01 *** p ≤ 0.001**** p ≤ 0.0005.

Article Snippet: SERCA1 , Rabbit , 1:1000 , Cell Signaling , #12293.

Techniques: Ex Vivo, Functional Assay, Muscles, Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Diaphragm Fatigue in SMNΔ7 Mice and Its Molecular Determinants: An Underestimated Issue

doi: 10.3390/ijms241914953

Figure Lengend Snippet: Oligonucleotide primers used for real-time PCR.

Article Snippet: SERCA1 , Rabbit , 1:1000 , Cell Signaling , #12293.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Diaphragm Fatigue in SMNΔ7 Mice and Its Molecular Determinants: An Underestimated Issue

doi: 10.3390/ijms241914953

Figure Lengend Snippet: Antibodies used for Western Blot Analysis.

Article Snippet: SERCA1 , Rabbit , 1:1000 , Cell Signaling , #12293.

Techniques: Western Blot

Journal: Skeletal Muscle

Article Title: Intracellular Ca 2+ -handling differs markedly between intact human muscle fibers and myotubes

doi: 10.1186/s13395-015-0050-x

Figure Lengend Snippet: The abundance of Ca 2+ -handling proteins differs markedly between human muscle fibers, myotubes and myoblasts. mRNA quantities of a RyR1 and DHPR and b SERCA1 and SERCA2 in human muscle fibers myotubes and myoblasts. Representative Western blot bands for c RyR and DHPR and d SERCA1 and SERCA2. Densitometry for the protein quantity of e RyR and DHPR and f SERCA1 and SERCA2 in human muscle fibers myotubes and myoblasts. Data are presented as the mean ± SEM. Asterisk denotes P < 0.05 relative to myoblasts, and number sign denotes P < 0.05 relative to myotubes. ( N/A ) not applicable due to protein quantity to low for reliable measurements

Article Snippet: Probes and primers (TaqMan) for SERCA1 (Hs01092295_m1), SERCA2 (Hs00544877_m1), RyR1 (Hs00166991_m1), α1s-DHPR (Hs00163885_m1), myogenin (Hs01072232_m1), MYH7 (Hs01110632_m1), MYH2 (HsHs0040042_m1), MYH1 (Hs00428600_m1), UBB (Hs00430290_m1), UBC (Hs00824723_m1), RPS18 (Hs01375212_g1), GAPDH (4352934E), 18S (4310893E), CTSB (Hs00157194_m1), and HK2 (Hs00606086_m1) were purchased from Applied Biosystems, Life Technologies.

Techniques: Western Blot

Journal: Skeletal Muscle

Article Title: Intracellular Ca 2+ -handling differs markedly between intact human muscle fibers and myotubes

doi: 10.1186/s13395-015-0050-x

Figure Lengend Snippet: The localization of Ca 2+ -handling proteins differs markedly between human muscle fibers and myotubes. a Representative immunofluorescent staining for RyR and DHPR in human muscle fibers ( top row ) and myotubes ( middle row ). b Magnification of the area indicated in a. c , d Fluorescent intensity (a.u.) for RyR ( black ) and DHPR ( red ) in a human muscle fiber ( left ) and myotube ( right ). e Representative immunofluorescent staining for SERCA1 and SERCA2 in human muscle fibers ( top row ) and myotubes ( middle row ). f Magnification of the area indicated in e . Scale bars indicate 10 μm. Primary antibody was omitted for the respective negative controls ( bottom row in a , b and e , f )

Article Snippet: Probes and primers (TaqMan) for SERCA1 (Hs01092295_m1), SERCA2 (Hs00544877_m1), RyR1 (Hs00166991_m1), α1s-DHPR (Hs00163885_m1), myogenin (Hs01072232_m1), MYH7 (Hs01110632_m1), MYH2 (HsHs0040042_m1), MYH1 (Hs00428600_m1), UBB (Hs00430290_m1), UBC (Hs00824723_m1), RPS18 (Hs01375212_g1), GAPDH (4352934E), 18S (4310893E), CTSB (Hs00157194_m1), and HK2 (Hs00606086_m1) were purchased from Applied Biosystems, Life Technologies.

Techniques: Staining

Journal: International Journal of Molecular Sciences

Article Title: Spatial and Functional Crosstalk between the Mitochondrial Na + -Ca 2+ Exchanger NCLX and the Sarcoplasmic Reticulum Ca 2+ Pump SERCA in Cardiomyocytes

doi: 10.3390/ijms23147948

Figure Lengend Snippet: Bimolecular fluorescence complementation assay for NCLX and NCLX-partner proteins—four isoforms of SERCA and NCX1—in HEK293 cells. ( A ) The complementary pairs of mKGN- and mKGC-fused NCLX and NCLX-partner proteins at N- or C-terminus are numbered as i–viii. ( B ) Representative images of i–viii complementary pairs for combinations NCLX–SERCA1, NCLX–SERCA2A, NCLX–SERCA3, and NCLX–NCX1. As a positive and a negative control, representative images of a mKGN-p65 and mKGC-p50 combination and a mKGN fused with NCLX N-terminus only are shown in the right, respectively. ( C ) Fluorescence intensity/area of positive cells, which were normalized with those of background cells. For combinations of NCLX and NCLX-partner proteins, data are expressed as mean ± s.e.m. of 8–37 cells. * p < 0.05, ** p < 0.01, compared with background. For positive control (PC) and negative control (NC), data are expressed as mean ± s.e.m. of 138 and 140 cells, respectively. ** p < 0.01, compared with negative control.

Article Snippet: The mouse open reading frame cDNA clones in pCMV6-Entry vector coding for SERCA1 (NM_007504), SERCA2A (NM_009722), SERCA2B (NM_001110140), and SERCA3 (NM_016745) were purchased from Origene Technologies.

Techniques: Bimolecular Fluorescence Complementation Assay, Negative Control, Fluorescence, Positive Control

Journal: The Journal of Physiology

Article Title: Slowed relaxation and preserved maximal force in soleus muscles of mice with targeted disruption of the Serca2 gene in skeletal muscle

doi: 10.1113/jphysiol.2011.211987

Figure Lengend Snippet: A, specificity of Serca2 gene disruption in skKO (Serca2flox/floxMlc-1fwt/cre) mice and control FF (Serca2flox/flox) mice. The presence of various gene alleles was analysed by PCR. Upper panel: loxP2, presence of the homozygous Serca2flox allele in all mice; wt, wt control PCR reaction. Middle panel: Cre, presence of Cre recombinase in the Mlc-1f gene locus; Fabpi 200 bp, autosomal gene internal PCR control. Lower panel: loxP1/P2, presence of the recombined and disrupted Serca2flox allele between loxP sites 1 and 2; Fabpi 500 bp, autosomal gene internal PCR control. The disrupted Serca2flox allele is present only in skKO skeletal muscle tissues (lower right). Tissues: LV, heart left ventricle; RV, heart right ventricle; atria, lung, kidney, liver, tibialis anterior, soleus, EDL, diaphragm, gastrocnemius and spleen. B, Serca2 mRNA quantification by RT-qPCR in skeletal muscle and other tissues as indicated from skKO and FF mice. Values are relative to abundance in FF soleus. C, summary quantification of relative SERCA2 protein abundance in selected tissues from skKO and FF mice. Values are normalized to FF. D, immuno-blots of SERCA2 protein in selected tissues from skKO and FF mice. E, immuno-blots of SERCA1 and SERCA3 protein in soleus muscle of skKO and FF mice. F, immuno-blots of total and phosphorylated phospholamban protein in soleus muscle of skKO and FF mice.

Article Snippet: Taqman gene expression assays (Applied Biosystems, Foster City, CA, USA) for Serca2 ( Atp2a2 , Mm01201431_m1), Serca1 ( Atp2a1 , Mm00476050_m1), Serca3 ( Atp2a3 , Mm00443897_m1 ), Ryr1 ( Ryr1 , Mm01175195_m1), Dhpr ( Cacna1s , Mm00489257_m1), Pmca1 ( Atp2b1 , Mm01245805_m1), Pmca4 ( Atp2b4 , Mm01285597_m1), Ncx1 ( Slc8a1 , Mm0441524_m1), Ncx2 ( Slc8a2 , Mm00455836_m1) Ncx3 ( Slc8a3 , Mm00475515_m1), Sarcalumenin ( Srl , Mm00614763_m1), Parvalbumin (Pvalb , Mm00443100_m1) and Gapdh ( Gapdh , Mm99999915_g1), were used.

Techniques: Disruption, Control, Quantitative RT-PCR, Quantitative Proteomics, Western Blot

Journal: The Journal of Physiology

Article Title: Slowed relaxation and preserved maximal force in soleus muscles of mice with targeted disruption of the Serca2 gene in skeletal muscle

doi: 10.1113/jphysiol.2011.211987

Figure Lengend Snippet: Staining of fixed cross-sections from skKO and FF soleus muscles as indicated. A–H, myosin heavy chain isoforms (MHCI, MHCII) and SERCA staining show that SERCA1 expression is restricted to fast-twitch fibres whereas SERCA2 expression is restricted to slow-twitch fibres. SERCA2 expression is completely absent in skKO slow-twitch fibres. 1 slow-twitch fibres positive for MHCI (A and B), negative for MHCII (E and F), negative for SERCA1 (G and H) and positive for SERCA2 (G), except for skKO muscle where SERCA2 is negative (H). 2 fast-twitch fibres negative for MHCI (A and B), positive for MHCII (C and D), positive for SERCA1 (E and F) and negative for SERCA2 (G and H). I and J, haematoxylin–eosin staining indicates similar basic structure and no sign of atrophy or degeneration of the skKO muscle fibres.

Article Snippet: Taqman gene expression assays (Applied Biosystems, Foster City, CA, USA) for Serca2 ( Atp2a2 , Mm01201431_m1), Serca1 ( Atp2a1 , Mm00476050_m1), Serca3 ( Atp2a3 , Mm00443897_m1 ), Ryr1 ( Ryr1 , Mm01175195_m1), Dhpr ( Cacna1s , Mm00489257_m1), Pmca1 ( Atp2b1 , Mm01245805_m1), Pmca4 ( Atp2b4 , Mm01285597_m1), Ncx1 ( Slc8a1 , Mm0441524_m1), Ncx2 ( Slc8a2 , Mm00455836_m1) Ncx3 ( Slc8a3 , Mm00475515_m1), Sarcalumenin ( Srl , Mm00614763_m1), Parvalbumin (Pvalb , Mm00443100_m1) and Gapdh ( Gapdh , Mm99999915_g1), were used.

Techniques: Staining, Muscles, Expressing

Journal: Poultry Science

Article Title: Improvement of eggshell quality by dietary N-carbamylglutamate supplementation in laying chickens

doi: 10.1016/j.psj.2020.04.004

Figure Lengend Snippet: Effects of NCG on expression of eggshell formation–related genes and proteins in the uterus. (A) Expression of Ca 2+ transport–related genes. (B) Expression of eggshell calcification–related genes (C) Expression of CALB1 and ATP2A1 proteins. Asterisks indicate significant differences (values are means ± SEM; ∗ P < 0.05, ∗∗ P < 0.01 and ∗∗∗ P < 0.001). Abbreviation: NCG, N-carbamylglutamate.

Article Snippet: After blocking with 5% skim milk, the PVDF membrane was incubated with corresponding primary antibodies including rabbit anti-CALB1 (1:1,000, ET1702-54) and rabbit anti-ATP2A1 (1:500, ER1803-34, Hangzhou HuaAn Biotechnology Co., Hangzhou, China) at 4°C overnight.

Techniques: Expressing

Journal: Poultry Science

Article Title: Improvement of eggshell quality by dietary N-carbamylglutamate supplementation in laying chickens

doi: 10.1016/j.psj.2020.04.004

Figure Lengend Snippet: Effects of NCG on expression of eggshell formation–related genes and proteins in the duodenum. (A) Expression of Ca 2+ transport–related genes. (B) Expression of eggshell calcification–related genes (C) Expression of CALB1 and ATP2A1 proteins. Asterisks indicate significant differences (values are means ± SEM; ∗ P < 0.05, ∗∗ P < 0.01 and ∗∗∗ P < 0.001). Abbreviation: NCG, N-carbamylglutamate.

Article Snippet: After blocking with 5% skim milk, the PVDF membrane was incubated with corresponding primary antibodies including rabbit anti-CALB1 (1:1,000, ET1702-54) and rabbit anti-ATP2A1 (1:500, ER1803-34, Hangzhou HuaAn Biotechnology Co., Hangzhou, China) at 4°C overnight.

Techniques: Expressing

Journal: Poultry Science

Article Title: Improvement of eggshell quality by dietary N-carbamylglutamate supplementation in laying chickens

doi: 10.1016/j.psj.2020.04.004

Figure Lengend Snippet: Effects of NCG on expression of eggshell formation–related genes and proteins in the kidney. (A) Expression of Ca 2+ transport–related genes. (B) Expression of eggshell calcification–related genes. (C) Expression of CALB1 and ATP2A1 proteins. Asterisks indicate significant differences (values are means ± SEM; ∗∗ P < 0.01 and ∗∗∗ P < 0.001). Abbreviation: NCG, N-carbamylglutamate.

Article Snippet: After blocking with 5% skim milk, the PVDF membrane was incubated with corresponding primary antibodies including rabbit anti-CALB1 (1:1,000, ET1702-54) and rabbit anti-ATP2A1 (1:500, ER1803-34, Hangzhou HuaAn Biotechnology Co., Hangzhou, China) at 4°C overnight.

Techniques: Expressing