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Proteintech bicinchoninic acid bca
Bicinchoninic Acid Bca, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/stim1+antibody/STIM1+Antibody/pmc13039107-268-6-31
Average 94 stars, based on 97 article reviews

bicinchoninic acid bca - by Bioz Stars, 2026-09

94/100 stars

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Article Title: STIM2 regulates NMDA receptor endocytosis that is induced by short-term NMDA receptor overactivation in cortical neurons
Article Snippet: Specific band for STIM1 and STIM2 was detected at approximately 90 kDa and 100 kDa, respectively, as observed by other authors using the same antibody from ProteinTech [ , – ].

Article Title: Transmembrane Protein Ttyh1 Maintains the Quiescence of Neural Stem Cells Through Ca 2+ /NFATc3 Signaling
Article Snippet: Membranes were blocked using 5% skim milk at room temperature for 1 h, and incubated at 4°C overnight with the addition of the primary antibodies of β-actin (1:10,000, Abcam), Ttyh1 (1:400, self-prepared , STIM1 (1:400, Proteintech, Chicago, IL, United States), Orai1 (1:400, Proteintech), NFATc3 (1:500, Proteintech), CALM (1:1,000, Abcam), CaMKII (1:1,000, Abcam), Cyclin D1 (1:5,000, Proteintech), p21 (1:1,000, Abcam).

Western Blot:

Article Title: Sodium Houttuyfonate Alleviates Monocrotaline-induced Pulmonary Hypertension by Regulating Canonical Transient Receptor Potential Channel Proteins.
Article Snippet: This is a PDF of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability.. This version will undergo additional copyediting, typesetting and review before it is published in its final form.. As such, this version is no longer the Accepted Manuscript, but it is not yet the definitive Version of Record; we are providing this early version to give early visibility of the article.

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(A) qPCR analysis of relative mRNA levels of indicated genes in BAT, epididymal white adipose tissue (eWAT) and inguinal white adipose tissue (iWAT). n= 2 mice per group (B) Left: Immunoblotting analysis of indicated proteins in total lysates from BAT, eWAT and iWAT from wild-type mice housed at RT. Right: Quantification analysis of the images shown in the left. n= 4 mice per group; One-way ANOVA, p< *0.05, *** p< <0.001. (C) qPCR analysis of relative mRNA levels of indicated genes in BAT, eWAT and iWAT. n= 4 per group; One-way ANOVA **p<0.005. (D, E) qPCR analysis of mRNA levels of indicated genes in iWAT (D) and eWAT (E) from mice exposed to 30°C, 22°C, and 4°C for 24hrs. In D, n= 3 (30°C); n=4 (22°C); n=3 (4°C); One-way ANOVA, * p<0.03. In E n= 4 (30°C); n= 4 (22°C); n=3 (4°C). (F) Left: Immunoblotting analysis of indicated protein in total lysates from BAT of mice housed at 30°C (thermoneutrality), 22°C (RT) or exposed to 4°C for 24 hrs. Right: Quantification analysis of images shown in the left. n= 3 per group (G) Upper panel: Representative bright field images of differentiated brown adipocytes (WT1 cells) control (scramble siRNA) or treated with STIM1/2 siRNA. Bottom panel: qPCR analysis of mRNA levels of indicated genes. n=3 per group. Unpaired Student’s t-test ***p<0.001. (H) Representative bright field images of differentiated primary brown adipocytes (after 7 days differentiation) from STIM1/2 flox and STIM1/2 AdpKO . (I, J) Immunostaining of STIM1 (green) in differentiated brown adipocytes in baseline and in cells treated with 1μM thapsigargin for 30 min in Ca 2+ free media (I) or 1μM or 10 μM Norepinephrine for 30 min in Ca 2+ free media (J). Nucleus are stained in blue.

Journal: bioRxiv

Article Title: Ca 2+ influx through ER-plasma membrane contacts is required for brown fat thermogenesis and metabolic health

doi: 10.64898/2026.03.20.712802

Figure Lengend Snippet: (A) qPCR analysis of relative mRNA levels of indicated genes in BAT, epididymal white adipose tissue (eWAT) and inguinal white adipose tissue (iWAT). n= 2 mice per group (B) Left: Immunoblotting analysis of indicated proteins in total lysates from BAT, eWAT and iWAT from wild-type mice housed at RT. Right: Quantification analysis of the images shown in the left. n= 4 mice per group; One-way ANOVA, p< *0.05, *** p< <0.001. (C) qPCR analysis of relative mRNA levels of indicated genes in BAT, eWAT and iWAT. n= 4 per group; One-way ANOVA **p<0.005. (D, E) qPCR analysis of mRNA levels of indicated genes in iWAT (D) and eWAT (E) from mice exposed to 30°C, 22°C, and 4°C for 24hrs. In D, n= 3 (30°C); n=4 (22°C); n=3 (4°C); One-way ANOVA, * p<0.03. In E n= 4 (30°C); n= 4 (22°C); n=3 (4°C). (F) Left: Immunoblotting analysis of indicated protein in total lysates from BAT of mice housed at 30°C (thermoneutrality), 22°C (RT) or exposed to 4°C for 24 hrs. Right: Quantification analysis of images shown in the left. n= 3 per group (G) Upper panel: Representative bright field images of differentiated brown adipocytes (WT1 cells) control (scramble siRNA) or treated with STIM1/2 siRNA. Bottom panel: qPCR analysis of mRNA levels of indicated genes. n=3 per group. Unpaired Student’s t-test ***p<0.001. (H) Representative bright field images of differentiated primary brown adipocytes (after 7 days differentiation) from STIM1/2 flox and STIM1/2 AdpKO . (I, J) Immunostaining of STIM1 (green) in differentiated brown adipocytes in baseline and in cells treated with 1μM thapsigargin for 30 min in Ca 2+ free media (I) or 1μM or 10 μM Norepinephrine for 30 min in Ca 2+ free media (J). Nucleus are stained in blue.

Article Snippet: STIM1 was stained with Rabbit anti-STIM1 (Cell Signaling, #4916) antibody overnight at 4°C, at a concentration of 1:200 in PBS.

Techniques: Western Blot, Control, Immunostaining, Staining

(A) Left: Immunoblotting analysis of indicated proteins in total lysates from BAT of mice maintained at 30°C (thermoneutrality), 22°C (RT) or exposed to 4°C (cold) for 24hrs. Right: Quantification analysis of images shown in A; n= 3 mice per group. One-way ANOVA with Tukey’s multiple comparisons test, *p<0.05, *** p<0.001. (B) mRNA levels of indicated genes in BAT from mice exposed to 30°C, 22°C, n= 4 and 4°C for 24hrs, n=3. One-way ANOVA with Tukey’s multiple comparisons test, * p<0.05, ** p<0.01, *** p<0.005. (C) Upper panel: Immunostaining of STIM1 (green) in differentiated brown adipocytes in baseline and in cells treated with 1μM thapsigargin (Tg) for 10 min in Ca 2+ free media; lipid droplets (LD) are stained in blue. Bottom: Quantification of the ratio of fluorescent intensity in the plasma membrane per intensity in the adjacent cytosolic area. n= 16 cells baseline, 10 cells Tg. Representative of 2 independent experiments; unpaired Student t-test ****p< <0.0001. (D) Left: Fluo-4 based cytosolic Ca 2+ measurements in differentiated primary brown adipocytes from STIM1/2 flox (control) and STIM1/2 AdpKO mice. Cells were first treated with 1μM Tg in Ca 2+ free media. Subsequently, 2mM Ca 2+ was added in the indicated time point. The graph shows a representative run of 2 independent experiments. n=15 cells for STIM1/2 flox and n=13 cells for STIM1/2 AdpKO . Right: Quantification of the cytosolic peak after Tg and Ca 2+ addition. Unpaired Student t-test *p<0.05. (E) Upper panel: Immunostaining of STIM1 (green) in differentiated brown adipocytes in baseline and in cells treated with 10μM norepinephrine (NE) for 30 min in Ca 2+ free media. Lipid droplets are stained in blue. Bottom: Quantification of the ratio of fluorescent intensity in the plasma membrane per intensity in the adjacent cytosolic area. n= 11 cells baseline, 8 cells NE. Representative of 3 independent experiments; Unpaired Student t-test ****p<0.0001. (F) Fluo-4-based cytosolic Ca 2+ measurements in differentiated primary brown adipocytes from STIM1/2 flox (control) and STIM1/2 AdpKO mice. Cells were first treated with 10μM NE in Ca 2+ free media. Subsequently, 2mM Ca 2+ was added in the indicated time point. n= 10 cells per group; Representative of 3 independent experiments. (G) Left: Fluo-4-based cytosolic Ca 2+ measurements in differentiated primary brown adipocytes from STIM1/2 flox and STIM1/2 AdpKO mice. Cells were treated with 10μM NE in the presence of 2mM Ca 2+ in the media. The graph shows a representative run of 2 independent experiments. n=23 for STIM1/2 flox , n=19 for STIM1/2 AdpKO . Right: Area under the curve of the graph shown on the left. Unpaired Student t- test *** p<0.001. (H) Rhod-2-based mitochondria Ca 2+ measurements in differentiated brown adipocytes transfected with scramble siRNA or with siRNA against STIM1/2. Cells were treated with 10μM NE in the presence of Ca 2+ in the media. Representative of 2 independent experiments.

Journal: bioRxiv

Article Title: Ca 2+ influx through ER-plasma membrane contacts is required for brown fat thermogenesis and metabolic health

doi: 10.64898/2026.03.20.712802

Figure Lengend Snippet: (A) Left: Immunoblotting analysis of indicated proteins in total lysates from BAT of mice maintained at 30°C (thermoneutrality), 22°C (RT) or exposed to 4°C (cold) for 24hrs. Right: Quantification analysis of images shown in A; n= 3 mice per group. One-way ANOVA with Tukey’s multiple comparisons test, *p<0.05, *** p<0.001. (B) mRNA levels of indicated genes in BAT from mice exposed to 30°C, 22°C, n= 4 and 4°C for 24hrs, n=3. One-way ANOVA with Tukey’s multiple comparisons test, * p<0.05, ** p<0.01, *** p<0.005. (C) Upper panel: Immunostaining of STIM1 (green) in differentiated brown adipocytes in baseline and in cells treated with 1μM thapsigargin (Tg) for 10 min in Ca 2+ free media; lipid droplets (LD) are stained in blue. Bottom: Quantification of the ratio of fluorescent intensity in the plasma membrane per intensity in the adjacent cytosolic area. n= 16 cells baseline, 10 cells Tg. Representative of 2 independent experiments; unpaired Student t-test ****p< <0.0001. (D) Left: Fluo-4 based cytosolic Ca 2+ measurements in differentiated primary brown adipocytes from STIM1/2 flox (control) and STIM1/2 AdpKO mice. Cells were first treated with 1μM Tg in Ca 2+ free media. Subsequently, 2mM Ca 2+ was added in the indicated time point. The graph shows a representative run of 2 independent experiments. n=15 cells for STIM1/2 flox and n=13 cells for STIM1/2 AdpKO . Right: Quantification of the cytosolic peak after Tg and Ca 2+ addition. Unpaired Student t-test *p<0.05. (E) Upper panel: Immunostaining of STIM1 (green) in differentiated brown adipocytes in baseline and in cells treated with 10μM norepinephrine (NE) for 30 min in Ca 2+ free media. Lipid droplets are stained in blue. Bottom: Quantification of the ratio of fluorescent intensity in the plasma membrane per intensity in the adjacent cytosolic area. n= 11 cells baseline, 8 cells NE. Representative of 3 independent experiments; Unpaired Student t-test ****p<0.0001. (F) Fluo-4-based cytosolic Ca 2+ measurements in differentiated primary brown adipocytes from STIM1/2 flox (control) and STIM1/2 AdpKO mice. Cells were first treated with 10μM NE in Ca 2+ free media. Subsequently, 2mM Ca 2+ was added in the indicated time point. n= 10 cells per group; Representative of 3 independent experiments. (G) Left: Fluo-4-based cytosolic Ca 2+ measurements in differentiated primary brown adipocytes from STIM1/2 flox and STIM1/2 AdpKO mice. Cells were treated with 10μM NE in the presence of 2mM Ca 2+ in the media. The graph shows a representative run of 2 independent experiments. n=23 for STIM1/2 flox , n=19 for STIM1/2 AdpKO . Right: Area under the curve of the graph shown on the left. Unpaired Student t- test *** p<0.001. (H) Rhod-2-based mitochondria Ca 2+ measurements in differentiated brown adipocytes transfected with scramble siRNA or with siRNA against STIM1/2. Cells were treated with 10μM NE in the presence of Ca 2+ in the media. Representative of 2 independent experiments.

Article Snippet: STIM1 was stained with Rabbit anti-STIM1 (Cell Signaling, #4916) antibody overnight at 4°C, at a concentration of 1:200 in PBS.

Techniques: Western Blot, Immunostaining, Staining, Clinical Proteomics, Membrane, Control, Transfection

(A) Schematic depicting the genetic strategy used to generate mice with adipose tissue specific deletion of STIM1/2 (STIM1/2 AdpKO ). AdpCre- adiponectin Cre. (B) Upper panel: Immunoblotting analysis of indicated proteins in total lysates from iWAT of STIM1/2 flox and STIM1/2 AdpCre mice. Bottom panel: Quantification analysis of images shown in upper panel; n= 5 mice per group. Unpaired Student’s t-test, **p< 0.01. (C) Upper panel: Immunoblotting analysis of indicated proteins in total lysates from eWAT of STIM1/2 flox and STIM1/2 AdpKO mice. Bottom panel: Quantification analysis of images shown in upper panel; n= 5 mice per group. Unpaired Student’s t-test, * p<0.05, **p<0.002. (D) Body weight gain curves of STIM1/2 flox and STIM1/2 AdpKO mice fed a chow diet. n = 6 mice per group. (E) Schematic depicting the strategy used to generate mice with brown adipose tissue specific deletion of STIM1/2. (F) Immunoblotting analysis of Cre recombinase in the indicated tissues from mice injected with AAV-Rec2-UPC1-Cre-miR122. (G) Upper panel: Immunoblotting analysis of indicated proteins in in total lysates from BAT, eWAT and iWAT. Bottom panel: Quantification analysis of images shown in upper panel; n= 3 mice per group. Unpaired Student’s t-test, **p< 0.01, *p<0.05. (H) Top: Representative H&E-stained sections of BAT from STIM1/2 flox and STIM1/2 AdpKO mice housed at RT. Bottom: Quantification analysis of area of LD and frequence distribution.

Journal: bioRxiv

Article Title: Ca 2+ influx through ER-plasma membrane contacts is required for brown fat thermogenesis and metabolic health

doi: 10.64898/2026.03.20.712802

Figure Lengend Snippet: (A) Schematic depicting the genetic strategy used to generate mice with adipose tissue specific deletion of STIM1/2 (STIM1/2 AdpKO ). AdpCre- adiponectin Cre. (B) Upper panel: Immunoblotting analysis of indicated proteins in total lysates from iWAT of STIM1/2 flox and STIM1/2 AdpCre mice. Bottom panel: Quantification analysis of images shown in upper panel; n= 5 mice per group. Unpaired Student’s t-test, **p< 0.01. (C) Upper panel: Immunoblotting analysis of indicated proteins in total lysates from eWAT of STIM1/2 flox and STIM1/2 AdpKO mice. Bottom panel: Quantification analysis of images shown in upper panel; n= 5 mice per group. Unpaired Student’s t-test, * p<0.05, **p<0.002. (D) Body weight gain curves of STIM1/2 flox and STIM1/2 AdpKO mice fed a chow diet. n = 6 mice per group. (E) Schematic depicting the strategy used to generate mice with brown adipose tissue specific deletion of STIM1/2. (F) Immunoblotting analysis of Cre recombinase in the indicated tissues from mice injected with AAV-Rec2-UPC1-Cre-miR122. (G) Upper panel: Immunoblotting analysis of indicated proteins in in total lysates from BAT, eWAT and iWAT. Bottom panel: Quantification analysis of images shown in upper panel; n= 3 mice per group. Unpaired Student’s t-test, **p< 0.01, *p<0.05. (H) Top: Representative H&E-stained sections of BAT from STIM1/2 flox and STIM1/2 AdpKO mice housed at RT. Bottom: Quantification analysis of area of LD and frequence distribution.

Article Snippet: STIM1 was stained with Rabbit anti-STIM1 (Cell Signaling, #4916) antibody overnight at 4°C, at a concentration of 1:200 in PBS.

Techniques: Western Blot, Injection, Staining

(A) Left: Immunoblotting analysis of indicated proteins in total lysates of BAT from STIM1/2 flox (controls) and STIM1/2 AdpKO . Right: Quantification of the images shown in the left. n=7 mice per group. Unpaired Student t-test, ****p<0.0001, *** p<0.005. (B, C) Body temperature over time of male (B) and female (C) mice exposed to 4°C in the absence of food. n =12 male mice per group and n =5 female mice per group. Two-way ANOVA with Šídák’s multiple comparisons test ***p<0.001,****p<0.0001. (D) Left: thermographic images from FLIR E5 infrared camera of STIM1/2 flox and STIM1/2 AdpKO mice exposed to 4°C for 4-6 hrs. Right: Quantification of images shown in the left. n= 6 mice per group. Unpaired Student t-test, *p<0.05. (E) Left: Norepinephrine-stimulated O2 consumption in anesthetized STIM1/2 flox and STIM1/2 AdpKO female mice. Right: Area under the curve of graph in the left. n= 6 mice STIM1/2 flox and n=5 mice STIM1/2 AdpKO . Unpaired Student’s t-test ** p<0.001. (F) Body temperature over time of male STIM1/2 flox mice expressing AAV-Rec2-UCP1-Null AAV-Rec2-UCP1-Cre recombinase exposed to 4°C in the absence of food. n=4 Null mice and n=3 Cre mice per group. Two-way ANOVA with Šídák’s multiple comparisons test * p<0.05, ** p< 0.01, *** p=.0001. (G) Left: Representative Hematoxylin and eosin (H&E) stained sections of BAT from STIM1/2 flox and STIM1/2 AdpKO mice exposed to cold for 6hrs. Right: Quantification analysis of lipid droplet (LD) number and size of images shown in the left. n=5 STIM1/2 flox , n=7 for STIM1/2 AdpKO . Unpaired Student t-test * p<0.05. (H) Left: Immunoblotting analysis of indicated proteins in total lysates of BAT from STIM1/2 flox and STIM1/2 AdpKO maintained at RT or exposed to cold (4°C) for 2hrs. Right: Quantification of the images shown in the left. n=3 per group. One-way ANOVA with Tukey’s multiple comparisons test. * p<0.05, ** p< 0.005.

Journal: bioRxiv

Article Title: Ca 2+ influx through ER-plasma membrane contacts is required for brown fat thermogenesis and metabolic health

doi: 10.64898/2026.03.20.712802

Figure Lengend Snippet: (A) Left: Immunoblotting analysis of indicated proteins in total lysates of BAT from STIM1/2 flox (controls) and STIM1/2 AdpKO . Right: Quantification of the images shown in the left. n=7 mice per group. Unpaired Student t-test, ****p<0.0001, *** p<0.005. (B, C) Body temperature over time of male (B) and female (C) mice exposed to 4°C in the absence of food. n =12 male mice per group and n =5 female mice per group. Two-way ANOVA with Šídák’s multiple comparisons test ***p<0.001,****p<0.0001. (D) Left: thermographic images from FLIR E5 infrared camera of STIM1/2 flox and STIM1/2 AdpKO mice exposed to 4°C for 4-6 hrs. Right: Quantification of images shown in the left. n= 6 mice per group. Unpaired Student t-test, *p<0.05. (E) Left: Norepinephrine-stimulated O2 consumption in anesthetized STIM1/2 flox and STIM1/2 AdpKO female mice. Right: Area under the curve of graph in the left. n= 6 mice STIM1/2 flox and n=5 mice STIM1/2 AdpKO . Unpaired Student’s t-test ** p<0.001. (F) Body temperature over time of male STIM1/2 flox mice expressing AAV-Rec2-UCP1-Null AAV-Rec2-UCP1-Cre recombinase exposed to 4°C in the absence of food. n=4 Null mice and n=3 Cre mice per group. Two-way ANOVA with Šídák’s multiple comparisons test * p<0.05, ** p< 0.01, *** p=.0001. (G) Left: Representative Hematoxylin and eosin (H&E) stained sections of BAT from STIM1/2 flox and STIM1/2 AdpKO mice exposed to cold for 6hrs. Right: Quantification analysis of lipid droplet (LD) number and size of images shown in the left. n=5 STIM1/2 flox , n=7 for STIM1/2 AdpKO . Unpaired Student t-test * p<0.05. (H) Left: Immunoblotting analysis of indicated proteins in total lysates of BAT from STIM1/2 flox and STIM1/2 AdpKO maintained at RT or exposed to cold (4°C) for 2hrs. Right: Quantification of the images shown in the left. n=3 per group. One-way ANOVA with Tukey’s multiple comparisons test. * p<0.05, ** p< 0.005.

Article Snippet: STIM1 was stained with Rabbit anti-STIM1 (Cell Signaling, #4916) antibody overnight at 4°C, at a concentration of 1:200 in PBS.

Techniques: Western Blot, Expressing, Staining

(A) Heat map of RNA-seq analysis of mRNA expression of BAT from STIM1/2 flox and STIM1/2 AdpKO at RT or exposed to cold (4°C) for 6hrs; n= 2 mice for STIM1/2 flox RT; n=3 mice for STIM1/2 AdpKO RT; n= 4 mice for STIM1/2 flox and STIM1/2 AdpKO cold (B) Volcano plots showing differentially expressed genes comparing STIM1/2 flox and STIM1/2 AdpKO exposed to cold. Red dots show the significant regulated genes (p<0.05), Unpaired Student’s t-test. (C, E) Ingenuity pathway analysis showing pathways predicted to be decreased (C) or increased (E) in STIM1/2 AdpKO compared to STIM1/2 flox exposed to cold (4°C). p-value was calculated using Benjamin-Hochberg analysis. (D, F) qPCR-based analysis of mRNA expression of indicated genes in BAT from STIM1/2 flox and STIM1/2 AdpKO at RT or exposed to 4°C for 6h; n= 2 mice STIM1/2 flox RT; n = 4 mice STIM1/2 AdpKO RT; n = 5 mice STIM1/2 flox cold; n= 8 mice STIM1/2 AdpKO cold. Unpaired Student’s t-test *p<0.05, **p<0.01, ***p<0.0005. (G) Relative activation of ATF6, XBP1s and PERK gene sets from RNA seq analysis shown in A. These analyses were performed as previously described . Differential activation of the ATF6, XBP1s, and PERK gene-sets was assessed by one-way ANOVA and significance of pairwise comparison confirmed by unpaired t-test. (H, I) SEM images of BAT tissue from STIM1/2 flox (H) and STIM1/2 AdpKO (I) mice exposed to cold (4°C) for 12hrs. White boxes highlight the ER/ aggregated membranes. (J) Segmentation and 3D reconstruction of FIB-SEM images of BAT from STIM1/2 flox and STIM1/2 AdpKO exposed to cold for 12hrs.

Journal: bioRxiv

Article Title: Ca 2+ influx through ER-plasma membrane contacts is required for brown fat thermogenesis and metabolic health

doi: 10.64898/2026.03.20.712802

Figure Lengend Snippet: (A) Heat map of RNA-seq analysis of mRNA expression of BAT from STIM1/2 flox and STIM1/2 AdpKO at RT or exposed to cold (4°C) for 6hrs; n= 2 mice for STIM1/2 flox RT; n=3 mice for STIM1/2 AdpKO RT; n= 4 mice for STIM1/2 flox and STIM1/2 AdpKO cold (B) Volcano plots showing differentially expressed genes comparing STIM1/2 flox and STIM1/2 AdpKO exposed to cold. Red dots show the significant regulated genes (p<0.05), Unpaired Student’s t-test. (C, E) Ingenuity pathway analysis showing pathways predicted to be decreased (C) or increased (E) in STIM1/2 AdpKO compared to STIM1/2 flox exposed to cold (4°C). p-value was calculated using Benjamin-Hochberg analysis. (D, F) qPCR-based analysis of mRNA expression of indicated genes in BAT from STIM1/2 flox and STIM1/2 AdpKO at RT or exposed to 4°C for 6h; n= 2 mice STIM1/2 flox RT; n = 4 mice STIM1/2 AdpKO RT; n = 5 mice STIM1/2 flox cold; n= 8 mice STIM1/2 AdpKO cold. Unpaired Student’s t-test *p<0.05, **p<0.01, ***p<0.0005. (G) Relative activation of ATF6, XBP1s and PERK gene sets from RNA seq analysis shown in A. These analyses were performed as previously described . Differential activation of the ATF6, XBP1s, and PERK gene-sets was assessed by one-way ANOVA and significance of pairwise comparison confirmed by unpaired t-test. (H, I) SEM images of BAT tissue from STIM1/2 flox (H) and STIM1/2 AdpKO (I) mice exposed to cold (4°C) for 12hrs. White boxes highlight the ER/ aggregated membranes. (J) Segmentation and 3D reconstruction of FIB-SEM images of BAT from STIM1/2 flox and STIM1/2 AdpKO exposed to cold for 12hrs.

Article Snippet: STIM1 was stained with Rabbit anti-STIM1 (Cell Signaling, #4916) antibody overnight at 4°C, at a concentration of 1:200 in PBS.

Techniques: RNA Sequencing, Expressing, Activation Assay, Comparison

(A, B) Representative TEM images of BAT from STIM1/2 flox (A) and STIM1/2 AdpKO (B) mice maintained at RT. (C, D) TEM images of BAT from STIM1/2 AdpKO exposed to cold highlighting the aggregated membranes from two different mice. (E) Representative TEM image of BAT from mice treated with tunicamycin (0.5mg/kg) for 6 hrs.

Journal: bioRxiv

Article Title: Ca 2+ influx through ER-plasma membrane contacts is required for brown fat thermogenesis and metabolic health

doi: 10.64898/2026.03.20.712802

Figure Lengend Snippet: (A, B) Representative TEM images of BAT from STIM1/2 flox (A) and STIM1/2 AdpKO (B) mice maintained at RT. (C, D) TEM images of BAT from STIM1/2 AdpKO exposed to cold highlighting the aggregated membranes from two different mice. (E) Representative TEM image of BAT from mice treated with tunicamycin (0.5mg/kg) for 6 hrs.

Article Snippet: STIM1 was stained with Rabbit anti-STIM1 (Cell Signaling, #4916) antibody overnight at 4°C, at a concentration of 1:200 in PBS.

Techniques:

(A, B) Reconstruction of individual mitochondria from FIB-SEM data of BAT from STIM1/2 flox (A) and STIM1/2 AdpKO mice (B). 3D reconstructions were performed using Zeiss Arivis Pro software. (C) Quantification of mitochondria volume from data shown in A and B. n = 886 and 445 mitochondria, respectively. Unpaired Student’s t-test ***p<0.001 (D) Quantification of mitochondria sphericity and frequence distribution from data shown in A and B. n = 624 mitochondria in STIM1/2 flox and n=337 mitochondria in STIM1/2 AdpKO . Unpaired Student’s t-test ****p<0.0001. (E) Mitochondria complexity index calculated according to . Unpaired Student’s t-test ****p<0.0001. (F) Confocal images from differentiated primary adipocytes derived from STIM1/2 flox and STIM1/2 AdpKO mice at baseline or treated with 10uM norepinephrine (NE) for 50 min. Mitochondria (green) was staining with OXPHOs antibody, LD (blue) and Nucleus (magenta). (G,H) Quantification of mitochondria aspect ratio (G) and mitochondria area (H) of images shown in F. STIM1/2 flox NT= 1689 mitochondria, NE=861 mitochondria; STIM1/2 AdpKO NT=1185 mitochondria, NE=1618 mitochondria from > 5 images. Representative of 3 independent experiments. One-way ANOVA with Tukey’s multiple comparisons test **** p<0.0001. (I) Upper panel: Immunoblotting analysis of indicated proteins in total lysates of BAT from STIM1/2 flox and STIM1/2 AdpKO mice maintained at RT or exposed to cold (4°C) for 2hrs. Bottom panel: Quantification analysis of images shown in the upper panel. n=3 mice per group for RT conditions and n=4 mice per group for cold conditions. One-way ANOVA with Tukey’s multiple comparisons test. **p< 0.005. (J) Seahorse based oxygen consumption rate of mitochondria isolated from STIM1/2 flox and STIM1/2 AdpKO exposed to cold (4°C) for 12 hrs. Arrows depicts the addition of 20mM Pyruvate + 20mM Malate; 2μg/mL Oligomycin, 9μM FCCP and 4 µM rotenone and 2 µM antimycin. n=10 wells per group. Representative of 3 independent experiments. (K) Quantification of basal and maximal respiration. Unpaired Student t-test ** p<0.001, **** p<0.0001.

Journal: bioRxiv

Article Title: Ca 2+ influx through ER-plasma membrane contacts is required for brown fat thermogenesis and metabolic health

doi: 10.64898/2026.03.20.712802

Figure Lengend Snippet: (A, B) Reconstruction of individual mitochondria from FIB-SEM data of BAT from STIM1/2 flox (A) and STIM1/2 AdpKO mice (B). 3D reconstructions were performed using Zeiss Arivis Pro software. (C) Quantification of mitochondria volume from data shown in A and B. n = 886 and 445 mitochondria, respectively. Unpaired Student’s t-test ***p<0.001 (D) Quantification of mitochondria sphericity and frequence distribution from data shown in A and B. n = 624 mitochondria in STIM1/2 flox and n=337 mitochondria in STIM1/2 AdpKO . Unpaired Student’s t-test ****p<0.0001. (E) Mitochondria complexity index calculated according to . Unpaired Student’s t-test ****p<0.0001. (F) Confocal images from differentiated primary adipocytes derived from STIM1/2 flox and STIM1/2 AdpKO mice at baseline or treated with 10uM norepinephrine (NE) for 50 min. Mitochondria (green) was staining with OXPHOs antibody, LD (blue) and Nucleus (magenta). (G,H) Quantification of mitochondria aspect ratio (G) and mitochondria area (H) of images shown in F. STIM1/2 flox NT= 1689 mitochondria, NE=861 mitochondria; STIM1/2 AdpKO NT=1185 mitochondria, NE=1618 mitochondria from > 5 images. Representative of 3 independent experiments. One-way ANOVA with Tukey’s multiple comparisons test **** p<0.0001. (I) Upper panel: Immunoblotting analysis of indicated proteins in total lysates of BAT from STIM1/2 flox and STIM1/2 AdpKO mice maintained at RT or exposed to cold (4°C) for 2hrs. Bottom panel: Quantification analysis of images shown in the upper panel. n=3 mice per group for RT conditions and n=4 mice per group for cold conditions. One-way ANOVA with Tukey’s multiple comparisons test. **p< 0.005. (J) Seahorse based oxygen consumption rate of mitochondria isolated from STIM1/2 flox and STIM1/2 AdpKO exposed to cold (4°C) for 12 hrs. Arrows depicts the addition of 20mM Pyruvate + 20mM Malate; 2μg/mL Oligomycin, 9μM FCCP and 4 µM rotenone and 2 µM antimycin. n=10 wells per group. Representative of 3 independent experiments. (K) Quantification of basal and maximal respiration. Unpaired Student t-test ** p<0.001, **** p<0.0001.

Article Snippet: STIM1 was stained with Rabbit anti-STIM1 (Cell Signaling, #4916) antibody overnight at 4°C, at a concentration of 1:200 in PBS.

Techniques: Software, Derivative Assay, Staining, Western Blot, Isolation

(A) Representative TEM images of BAT from STIM1/2 flox and STIM1/2 AdpKO exposed to cold (4°C). (B) Left: Immunoblotting analysis of indicated proteins in isolated mitochondria from BAT of STIM1/2 flox and STIM1/2 AdpCre mice exposed to cold for 12hrs. Right: Quantification analysis of images shown in left panel; n= 3 mice per group. (C) Quantification strategy for images shown in . (D) Left: Immunoblotting analysis of indicated proteins in total lysates of BAT from STIM1/2 flox and STIM1/2 AdpKO mice maintained at RT or exposed to cold (4°C) for 2hrs. Right: Quantification analysis of images shown in the upper panel. n=3 mice per group for RT conditions and n=4 mice per group for cold conditions. One-way ANOVA with Tukey’s multiple comparisons test.

Journal: bioRxiv

Article Title: Ca 2+ influx through ER-plasma membrane contacts is required for brown fat thermogenesis and metabolic health

doi: 10.64898/2026.03.20.712802

Figure Lengend Snippet: (A) Representative TEM images of BAT from STIM1/2 flox and STIM1/2 AdpKO exposed to cold (4°C). (B) Left: Immunoblotting analysis of indicated proteins in isolated mitochondria from BAT of STIM1/2 flox and STIM1/2 AdpCre mice exposed to cold for 12hrs. Right: Quantification analysis of images shown in left panel; n= 3 mice per group. (C) Quantification strategy for images shown in . (D) Left: Immunoblotting analysis of indicated proteins in total lysates of BAT from STIM1/2 flox and STIM1/2 AdpKO mice maintained at RT or exposed to cold (4°C) for 2hrs. Right: Quantification analysis of images shown in the upper panel. n=3 mice per group for RT conditions and n=4 mice per group for cold conditions. One-way ANOVA with Tukey’s multiple comparisons test.

Article Snippet: STIM1 was stained with Rabbit anti-STIM1 (Cell Signaling, #4916) antibody overnight at 4°C, at a concentration of 1:200 in PBS.

Techniques: Western Blot, Isolation

(A) Left: Immunoblotting analysis of indicated proteins in total lysates from BAT of mice fed chow diet of high fat diet for 16 weeks. Right: Quantification analysis of the images shown in A; n= 6 mice for chow diet, n=6 mice for HFD group. Unpaired Student’s t-test, *p<0.05 (B) Body weight gain of STIM1/2 flox and STIM1/2 AdpKO mice fed a HFD over the indicated time. n=16 STIM1/2 flox mice, n=10 STIM1/2 AdpKO mice. (C) Insulin tolerance test, n =15 STIM1/2 flox mice, n=10 STIM1/2 AdpKO mice fed a HFD for 10 weeks. Two-way ANOVA **p < 0.01. (D) Oral glucose tolerance test, n=16 STIM1/2 flox mice, n=10 STIM1/2 AdpKO mice fed a HFD for 11 weeks (E) Insulin levels during the oral glucose tolerance test shown in D, Two-way ANOVA *p <0.05. (F) Representative hematoxylin and eosin-stained histology sections of BAT from STIM1/2 flox and STIM1/2 AdpKO mice fed HFD for 14 weeks. (G) Schematic depicting protocol of high fat diet (HFD) feeding and AAV-Rec2-UCP1-Null or - AAV-Rec2-UCP1-Cre injection. (H) Left: Immunoblotting analysis of indicated proteins in total lysates from BAT of mice fed HFD infected with AAV-Rec2-UCP1-Null or - AAV-Rec2-UCP1-Cre. Right: Quantification analysis of the images shown in A; n= 5 mice per group. Unpaired Student’s t-test, **p<0.01. (I) Insulin tolerance test, n= 4 Null mice and n=5 Cre mice; Two-way ANOVA *p <0.05. (J) Left: Oral glucose tolerance test, n =5 mice in each group. Two-way ANOVA *p <0.05. Right: Area under the curve from the graph shown on the left. Unpaired Student’s t-test *p<0.05

Journal: bioRxiv

Article Title: Ca 2+ influx through ER-plasma membrane contacts is required for brown fat thermogenesis and metabolic health

doi: 10.64898/2026.03.20.712802

Figure Lengend Snippet: (A) Left: Immunoblotting analysis of indicated proteins in total lysates from BAT of mice fed chow diet of high fat diet for 16 weeks. Right: Quantification analysis of the images shown in A; n= 6 mice for chow diet, n=6 mice for HFD group. Unpaired Student’s t-test, *p<0.05 (B) Body weight gain of STIM1/2 flox and STIM1/2 AdpKO mice fed a HFD over the indicated time. n=16 STIM1/2 flox mice, n=10 STIM1/2 AdpKO mice. (C) Insulin tolerance test, n =15 STIM1/2 flox mice, n=10 STIM1/2 AdpKO mice fed a HFD for 10 weeks. Two-way ANOVA **p < 0.01. (D) Oral glucose tolerance test, n=16 STIM1/2 flox mice, n=10 STIM1/2 AdpKO mice fed a HFD for 11 weeks (E) Insulin levels during the oral glucose tolerance test shown in D, Two-way ANOVA *p <0.05. (F) Representative hematoxylin and eosin-stained histology sections of BAT from STIM1/2 flox and STIM1/2 AdpKO mice fed HFD for 14 weeks. (G) Schematic depicting protocol of high fat diet (HFD) feeding and AAV-Rec2-UCP1-Null or - AAV-Rec2-UCP1-Cre injection. (H) Left: Immunoblotting analysis of indicated proteins in total lysates from BAT of mice fed HFD infected with AAV-Rec2-UCP1-Null or - AAV-Rec2-UCP1-Cre. Right: Quantification analysis of the images shown in A; n= 5 mice per group. Unpaired Student’s t-test, **p<0.01. (I) Insulin tolerance test, n= 4 Null mice and n=5 Cre mice; Two-way ANOVA *p <0.05. (J) Left: Oral glucose tolerance test, n =5 mice in each group. Two-way ANOVA *p <0.05. Right: Area under the curve from the graph shown on the left. Unpaired Student’s t-test *p<0.05

Article Snippet: STIM1 was stained with Rabbit anti-STIM1 (Cell Signaling, #4916) antibody overnight at 4°C, at a concentration of 1:200 in PBS.

Techniques: Western Blot, Staining, Injection, Infection

Calcium image analysis in PQ-injured A549 cells compared to normal cells ( A ) and PQ-injured A549 cells w/wo lysine supplementation ( B ) with thapsigargin (TG) or ionomycin (IONO) stimulation. Mean ± SEM. C NFAT luciferase expression in A549 cells treated w/wo 800 μM PQ, together with 0-, 1-, 2.5-, or 5-fold lysine as in the culture medium as indicated, for 24 h. Mean ± SD, *** P < 0.001; Two-Way ANOVA. n = 3. NS not significant. D WB analysis of E-Cadherin, ZO-1, EPCAM and acetyl-α-Tubulin in PQ-injured A549 cells treated w/wo 5 μM SKF-96365 (SKF). E Co-IP analysis of STIM1 association with ORAI1 or TRPC1 in PQ-injured A549 cells with indicated concentration, together w/wo 5 mM lysine. F Immunofluorescence analysis of Myc and ARL13B in PQ-injured TRPC1-Myc stably expressed A549 cells w/wo lysine supplementation. Scale bar, 10 μm.

Journal: Cell Death Discovery

Article Title: Lysine attenuates acute lung injury by restoring α-tubulin acetylation and ciliary activity

doi: 10.1038/s41420-026-03025-x

Figure Lengend Snippet: Calcium image analysis in PQ-injured A549 cells compared to normal cells ( A ) and PQ-injured A549 cells w/wo lysine supplementation ( B ) with thapsigargin (TG) or ionomycin (IONO) stimulation. Mean ± SEM. C NFAT luciferase expression in A549 cells treated w/wo 800 μM PQ, together with 0-, 1-, 2.5-, or 5-fold lysine as in the culture medium as indicated, for 24 h. Mean ± SD, *** P < 0.001; Two-Way ANOVA. n = 3. NS not significant. D WB analysis of E-Cadherin, ZO-1, EPCAM and acetyl-α-Tubulin in PQ-injured A549 cells treated w/wo 5 μM SKF-96365 (SKF). E Co-IP analysis of STIM1 association with ORAI1 or TRPC1 in PQ-injured A549 cells with indicated concentration, together w/wo 5 mM lysine. F Immunofluorescence analysis of Myc and ARL13B in PQ-injured TRPC1-Myc stably expressed A549 cells w/wo lysine supplementation. Scale bar, 10 μm.

Article Snippet: The lysates were quantified by the Bicinchoninic acid (BCA) method and incubated with STIM1 antibody (Cell Signaling Technology, 5668, 1 μl/sample) overnight, followed by incubation with 20 μl protein A/G beads (Proteintech, PR40025) for another 3 h at 4 °C.

Techniques: Luciferase, Expressing, Co-Immunoprecipitation Assay, Concentration Assay, Immunofluorescence, Stable Transfection

A GSEA analysis uncovered top-ranked molecular functions enhanced in PQ-injured lung tissues with lysine supplementation compared to the untreated group. B KEGG enrichment of the top 20 upregulated pathways in PQ-injured lung tissues with lysine supplementation compared to the untreated group. C Quantification of NADP and NADPH (left) or NAD and NADH (right) in PQ-injured A549 cells w/wo lysine supplementation. D ELISA analysis of acetyl coenzyme A (Acetyl-CoA) in PQ-injured A549 cells w/wo lysine supplementation. E WB analysis of E-Cadherin and acetyl-α-Tubulin in PQ-injured A549 cells treated w/wo 1 mM sodium acetate (SA). F WB analysis of ZO-1, E-Cadherin, acetyl-α-Tubulin and EPCAM in PQ-injured A549 cells w/wo lysine supplementation, together w/wo 0.5 μM GM-90257 (GM). G Calcium image analysis in PQ-injured A549 cells w/wo lysine supplementation, together w/wo 0.5 μM GM-90257 treatment. The cells were transiently stimulated with thapsigargin (TG) or ionomycin (IONO). Mean ± SEM. H Co-IP analysis of STIM1 association with TRPC1 in PQ-injured A549 cells w/wo lysine supplementation, together w/wo GM-90257 treatment. Immunofluorescence analysis of ARL13B, SFTPC, and HOPX in A549 cells ( I ) or normal lung tissues ( J ). Scale bar, 10 μm. Mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001; Two-Way ANOVA. NS not significant.

Journal: Cell Death Discovery

Article Title: Lysine attenuates acute lung injury by restoring α-tubulin acetylation and ciliary activity

doi: 10.1038/s41420-026-03025-x

Figure Lengend Snippet: A GSEA analysis uncovered top-ranked molecular functions enhanced in PQ-injured lung tissues with lysine supplementation compared to the untreated group. B KEGG enrichment of the top 20 upregulated pathways in PQ-injured lung tissues with lysine supplementation compared to the untreated group. C Quantification of NADP and NADPH (left) or NAD and NADH (right) in PQ-injured A549 cells w/wo lysine supplementation. D ELISA analysis of acetyl coenzyme A (Acetyl-CoA) in PQ-injured A549 cells w/wo lysine supplementation. E WB analysis of E-Cadherin and acetyl-α-Tubulin in PQ-injured A549 cells treated w/wo 1 mM sodium acetate (SA). F WB analysis of ZO-1, E-Cadherin, acetyl-α-Tubulin and EPCAM in PQ-injured A549 cells w/wo lysine supplementation, together w/wo 0.5 μM GM-90257 (GM). G Calcium image analysis in PQ-injured A549 cells w/wo lysine supplementation, together w/wo 0.5 μM GM-90257 treatment. The cells were transiently stimulated with thapsigargin (TG) or ionomycin (IONO). Mean ± SEM. H Co-IP analysis of STIM1 association with TRPC1 in PQ-injured A549 cells w/wo lysine supplementation, together w/wo GM-90257 treatment. Immunofluorescence analysis of ARL13B, SFTPC, and HOPX in A549 cells ( I ) or normal lung tissues ( J ). Scale bar, 10 μm. Mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001; Two-Way ANOVA. NS not significant.

Techniques: Enzyme-linked Immunosorbent Assay, Co-Immunoprecipitation Assay, Immunofluorescence

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