Journal: Physiological reports

Article Title: Environment-induced heat stress causes ventricular-dependent biochemical changes in the heart in female pigs.

doi: 10.14814/phy2.70414

Figure Lengend Snippet: FIGURE 4 Environment-induced heat stress altered calcium regulatory proteins in LV. In LV (a) environment-induced heat stress increased the relative protein abundance of the PLB/SERCA2a complex, STIM1, CaMKII, p-PKC pan, and p-PKC alpha/beta II compared to TN; while the monomeric and oligomeric forms of PLB, p-CaMKII, and p-PLB were similar between groups. In RV (b) relative protein abundance of the oligomeric form of PLB and p-PKC alpha/beta II increased by EIHS compared to TN; whereas the monomeric form of PLB, PLB/SERCA2a complex, STIM1, CaMKII, p-CaMKII, p-PLB, and p-PKC pan were similar between groups. Treatments: EIHS, environment-induced heat stress; TN, thermoneutral. Results are expressed as means ± standard deviation. * indicates p < 0.05. (a) LV: Monomer, oligomer, SERCA2a complex, CaMKII, p-CaMKII, p-PLB, p-PKC pan, and p-PKC alpha/beta II n = 7/group, one outlier from EIHS CaMKII and p-CaMKII and one outlier removed from TN p = PKC pan and p-PKC alpha/beta II; STIM1 n = 6/group, one outlier removed from EIHS. (b) RV: Monomer, oligomer, SERCA2a complex, CaMKII, p-CaMKII, p-PLB, n = 7/group, one outlier removed from TN oligomer; STIM1 n = 8/group, one outlier removed EIHS; p-PKC pan and p-PKC alpha/beta II TN n = 6/group, EIHS n = 8/group, one outlier removed from EIHS p-PKC alpha/beta II.

Article Snippet: Antibody Company/product No. Primary dilution Secondary dilution Calmodulin (CaM) Abcam, ab45689 1:1000 1:2000 Calmodulin- dependent protein kinase II (CaMKII) Santa Cruz Technologies (SC), #5306 1:500 1:1000 Phophorylated- Calmodulin- dependent protein kinase II (p- CaMKII) (Thr286) Cell Signaling Techology (CST), #12716 1:500 1:1000 Calpain 1 Large Subunit (Mu- Type CST, #2556 1:1000 1:2000 Calpain 2 Large Subunit (M- Type) CST, #2539 1:1000 1:2000 Calpastatin CST, #4146 1:1000 1:2000 Troponin T Millipore Sigma, #T6277 1:1000 1:2000 Calsequestrin Invitrogen, #VIIID12 1:500 1:1000 Calumenin SC, #271357 1:1000 1:2000 Junctophilin2 SC, #377086 1:1000 1:2000 Muscle atrophy F- box protein (MAFbx/Atrogin- 1) SC, #166806 1:1000 1:2000 Muscle RING finger protein 1 (MuRF- 1) SC, #398608 1:1000 1:2000 ORAI1 ProteinTech, #66223–1 1:1000 1:3000 Plasma membrane calcium ATPase (PMCA) SC, #211917 1:1000 1:2000 mouse Phospholamban (PLB) Novus, #NBP2- 19807 1:3000 (5% milk) 1:3000 (5% milk) Phosphorylated- Phospholamban (S16/Thr17) CST, #8496 1:1000 1:2000 Phosphorylated Protein Kinase C (p- PKC) Novus Biologicals, #BP2- 19807 1:3000 (5% milk) 1:3000 (5% milk) Phosphorylated Protein Kinase C (p- PKC) CST, #9580 1:1000 (5% milk) 1:4000 (5% milk) Ryanodine Receptor 2 (RyR2) Invitrogen, #MA3- 916 1:1000 1:2000 Phospho- Ryanodine Receptor (p- RyR2) Invitrogen, #PA5- 36758 1:1000 1:2000 Sarco/endoplasmic Reticulum Calcium ATPase (ATP2A2/ SERCA2) CST, #4388 1:1000 (5% milk) 1:4000 (5% milk) Sodium/Calcium exchanger 1 (NXC1) ProteinTech, #28447 1:1000 1:2000 Stromal interaction molecule 1 (STIM1) CST, #4916 1:1000 1:2000 Ubiquitin CST, #3933 1:1000 1:2000 Voltage dependent anion channel 2 (VDAC2) CST, #9412 1:1000 1:2000 nloaded from https://physoc.onlinelibrary.w iley.com /doi/10.14814/phy2.70414, W iley O nline L ibrary on [10/06/2025].

Techniques: Quantitative Proteomics, Standard Deviation

Journal: Physiological Reports

Article Title: Environment‐induced heat stress causes ventricular‐dependent biochemical changes in the heart in female pigs

doi: 10.14814/phy2.70414

Figure Lengend Snippet: Environment‐induced heat stress altered calcium regulatory proteins in LV. In LV (a) environment‐induced heat stress increased the relative protein abundance of the PLB/SERCA2a complex, STIM1, CaMKII, p‐PKC pan, and p‐PKC alpha/beta II compared to TN; while the monomeric and oligomeric forms of PLB, p‐CaMKII, and p‐PLB were similar between groups. In RV (b) relative protein abundance of the oligomeric form of PLB and p‐PKC alpha/beta II increased by EIHS compared to TN; whereas the monomeric form of PLB, PLB/SERCA2a complex, STIM1, CaMKII, p‐CaMKII, p‐PLB, and p‐PKC pan were similar between groups. Treatments: EIHS, environment‐induced heat stress; TN, thermoneutral. Results are expressed as means ± standard deviation. * indicates p < 0.05. (a) LV: Monomer, oligomer, SERCA2a complex, CaMKII, p‐CaMKII, p‐PLB, p‐PKC pan, and p‐PKC alpha/beta II n = 7/group, one outlier from EIHS CaMKII and p‐CaMKII and one outlier removed from TN p = PKC pan and p‐PKC alpha/beta II; STIM1 n = 6/group, one outlier removed from EIHS. (b) RV: Monomer, oligomer, SERCA2a complex, CaMKII, p‐CaMKII, p‐PLB, n = 7/group, one outlier removed from TN oligomer; STIM1 n = 8/group, one outlier removed EIHS; p‐PKC pan and p‐PKC alpha/beta II TN n = 6/group, EIHS n = 8/group, one outlier removed from EIHS p‐PKC alpha/beta II.

Article Snippet: Stromal interaction molecule 1 (STIM1) , CST, #4916 , 1:1000 , 1:2000.

Techniques: Quantitative Proteomics, Standard Deviation

Journal: Scientific Reports

Article Title: The miR-641-STIM1 and SATB1 axes play important roles in the regulation of the Th17/Treg balance in ITP

doi: 10.1038/s41598-024-61660-9

Figure Lengend Snippet: ITP Patients and Normal Candidates.

Article Snippet: The membranes were incubated with an anti-SATB1 antibody ([EPR3951] (ab109122, Abcam), an anti-Stromal interaction molecule 1 antibody (STIM1) ([EPR3414] (ab108994), an anti-β-Actin monoclonal antibody (66009-1-Ig, Proteintech), and appropriate horseradish peroxidase (HRP)-linked anti-rabbit/mouse IgG secondary antibodies (SA00001-1, SA00001-2, Proteintech) according to the primary antibody species.

Techniques: Biomarker Discovery

Journal: Scientific Reports

Article Title: The miR-641-STIM1 and SATB1 axes play important roles in the regulation of the Th17/Treg balance in ITP

doi: 10.1038/s41598-024-61660-9

Figure Lengend Snippet: ( A ) qRT-PCR analyzed miR641 expression in normal control (NC, blue, n = 5) and ITP patient (ITP, Red, n = 7), * p < 0.05. ( B ) qRT-PCR analyzed SATB1 expression in normal control (NC, blue, n = 5) and ITP patients (ITP, Red, n = 5), *** p < 0.001. ( C ) qRT-PCR analyzed STIM1 expression in normal control (NC, blue, n = 5) and ITP patients (ITP, Red, n = 5), *** p < 0.001. All groups underwent three technical replicates for qRT-PCR.

Article Snippet: The membranes were incubated with an anti-SATB1 antibody ([EPR3951] (ab109122, Abcam), an anti-Stromal interaction molecule 1 antibody (STIM1) ([EPR3414] (ab108994), an anti-β-Actin monoclonal antibody (66009-1-Ig, Proteintech), and appropriate horseradish peroxidase (HRP)-linked anti-rabbit/mouse IgG secondary antibodies (SA00001-1, SA00001-2, Proteintech) according to the primary antibody species.

Techniques: Quantitative RT-PCR, Expressing, Control

Journal: Scientific Reports

Article Title: The miR-641-STIM1 and SATB1 axes play important roles in the regulation of the Th17/Treg balance in ITP

doi: 10.1038/s41598-024-61660-9

Figure Lengend Snippet: ( A ) Luciferase Assay analyzed the relative Luciferase Activity (NC, blue column; 3’UTR Vector, Red column). NS. non significant. ( B ) Luciferase Assay analyzed the relative Luciferase Activity (NC, blue column; h-SATB1-WT, Red column). ( C ) Luciferase Assay analyzed the relative Luciferase Activity (NC, blue column; h-STIM1-WT, Red column). All groups underwent three technical replicates. D. qRT-PCR analyzed miR641 expression in 293 T cells after miR641 over-expression (lentivirus transfection), (Vecter, blue column; miR641 over expression, Red column), * p < 0.05 . E. qRT-PCR analyzed SATB1 expression in 293 T cells after miR641 over-expression(lentivirus transfection), (Vecter, blue column; miR641 over -expression, Red column), * p < 0.05. F. qRT-PCR analyzed STIM1 expression in 293 T cells after miR641 over-expression (lentivirus transfection), (Vecter, blue column; miR641 over- expression, Red column), * p < 0.05 . G. Western blotting analyzed the expression SATB1 and STIM1 encoded proteins after miR641 over-expression (lentivirus transfection) in 293 T cells (Vector Left; miR641 over-expression Right). All groups underwent three technical replicates.

Article Snippet: The membranes were incubated with an anti-SATB1 antibody ([EPR3951] (ab109122, Abcam), an anti-Stromal interaction molecule 1 antibody (STIM1) ([EPR3414] (ab108994), an anti-β-Actin monoclonal antibody (66009-1-Ig, Proteintech), and appropriate horseradish peroxidase (HRP)-linked anti-rabbit/mouse IgG secondary antibodies (SA00001-1, SA00001-2, Proteintech) according to the primary antibody species.

Techniques: Luciferase, Activity Assay, Plasmid Preparation, Quantitative RT-PCR, Expressing, Over Expression, Transfection, Western Blot

Journal: Scientific Reports

Article Title: The miR-641-STIM1 and SATB1 axes play important roles in the regulation of the Th17/Treg balance in ITP

doi: 10.1038/s41598-024-61660-9

Figure Lengend Snippet: ( A ) qRT-PCR analyzed miR641 expression after miR641 over-expression (lentivirus transfection) in primary T cells from 3 normal candidates. (Vector, blue column; miR641 over-expression, Red Column), * p < 0.05. B. qRT-PCR analyzed SATB1 expression after miR641 over-expression (lentivirus transfection) in primary T cells from 3 normal candidates (Vector, blue column; miR641 over-expression, Red Column), * p < 0.05. ( C ) qRT-PCR analyzed STIM1 expression after miR641 over-expression (lentivirus transfection) in primary T cells from 3 normal candidates. (Vector, blue column; miR641 over-expression, Red Column), * p < 0.05. ( D , E ) FACS analyzed the Th17 cells surface markers CD4 and IL-17A expression after miR641 over-expression (lentivirus transfection) in primary T cells from 3 normal candidates. ( F ) Statistic analysis of FACS results: CD4 + IL17A + Th17 cells were up-regulated in miR641 over-expression primary T cells from normal candidates (Red Column) comparing with vector only group (Blue Column). ( G , H ) FACS analyzed the Treg cells surface markers CD25 and Foxp3 expression after miR641 over-expression (lentivirus transfection) in primary T cells from 3 normal candidates. ( I ) Statistic analysis of FACS results: CD25 + Foxp3 + cells were up-regulated in miR641 over-expression primary T cells from 3 normal candidates (Red Column) comparing with vector only group (Blue Column).

Article Snippet: The membranes were incubated with an anti-SATB1 antibody ([EPR3951] (ab109122, Abcam), an anti-Stromal interaction molecule 1 antibody (STIM1) ([EPR3414] (ab108994), an anti-β-Actin monoclonal antibody (66009-1-Ig, Proteintech), and appropriate horseradish peroxidase (HRP)-linked anti-rabbit/mouse IgG secondary antibodies (SA00001-1, SA00001-2, Proteintech) according to the primary antibody species.

Techniques: Quantitative RT-PCR, Expressing, Over Expression, Transfection, Plasmid Preparation

Journal: Scientific Reports

Article Title: The miR-641-STIM1 and SATB1 axes play important roles in the regulation of the Th17/Treg balance in ITP

doi: 10.1038/s41598-024-61660-9

Figure Lengend Snippet: ( A ) qRT-PCR analyzed miR641 expression in miR641sponge transfected primary T cells from 3 ITP patients. (Sponge Control, blue column; miR641 sponge, Red Column), * p < 0.05. ( B ) qRT-PCR analyzed SATB1 expression in miR641sponge transfected primary T cells from 3 ITP patients. (Sponge Control, blue column; miR641 sponge, Red Column), * p < 0.05. ( C ) qRT-PCR analyzed STIM1 expression in miR641sponge transfected primary T cells from 3 ITP patients. (Sponge Control, blue column; miR641 sponge, Red Column), * p < 0.05. ( D , E ) FACS analyzed the Th17 cells surface markers CD4 and IL-17A expression in miR641 sponge transfected primary T cells from 3 ITP patients. ( F ) Statistic analysis of FACS results: CD4 + IL17A + Th17 cells were down-regulated in miR641sponge transfected primary T cells from 3 ITP patients (Red Column) comparing with Sponge Control group (Blue Column). ( G , H ) FACS analyzed the Treg cells surface markers CD25 and Foxp3 expression in miR641 sponge transfected primary T cells from 3 ITP patients. ( I ) Statistic analysis of FACS results: CD25 + Foxp3 + Treg cells were up-regulated in miR641sponge transfected primary T cells fromITP patients (Red Column) comparing with Sponge Control group (Blue Column).

Article Snippet: The membranes were incubated with an anti-SATB1 antibody ([EPR3951] (ab109122, Abcam), an anti-Stromal interaction molecule 1 antibody (STIM1) ([EPR3414] (ab108994), an anti-β-Actin monoclonal antibody (66009-1-Ig, Proteintech), and appropriate horseradish peroxidase (HRP)-linked anti-rabbit/mouse IgG secondary antibodies (SA00001-1, SA00001-2, Proteintech) according to the primary antibody species.

Techniques: Quantitative RT-PCR, Expressing, Transfection, Control

Journal: Scientific Reports

Article Title: The miR-641-STIM1 and SATB1 axes play important roles in the regulation of the Th17/Treg balance in ITP

doi: 10.1038/s41598-024-61660-9

Figure Lengend Snippet: ( A ) Schema of in vivo experiments. ( B ) qRT-PCR analyzed miR641 expression in the peripheral blood from MOCK group (dark blue column), ITP group (Red column) and AntagomiR641group (light blue column). ( C ) qRT-PCR analyzed SATB1 expression in the peripheral blood from MOCK group (dark blue column), ITP group (Red column) and AntagomiR641group (light blue column). ( D ) qRT-PCR analyzed STIM1 expression in the peripheral blood from MOCK group (dark blue column), ITP group (Red column) and AntagomiR641group (light blue column). qRT-PCR analysis underwent biological replication (n = 3).

Article Snippet: The membranes were incubated with an anti-SATB1 antibody ([EPR3951] (ab109122, Abcam), an anti-Stromal interaction molecule 1 antibody (STIM1) ([EPR3414] (ab108994), an anti-β-Actin monoclonal antibody (66009-1-Ig, Proteintech), and appropriate horseradish peroxidase (HRP)-linked anti-rabbit/mouse IgG secondary antibodies (SA00001-1, SA00001-2, Proteintech) according to the primary antibody species.

Techniques: In Vivo, Quantitative RT-PCR, Expressing