anti orai1  (Novus Biologicals)

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: Specific antibodies were used for STIM1 (Cell Signaling Technology, 5668, 1:3000), ORAI1 (Santa Cruz, sc-377281, 1:500), and TRPC1 (Proteintech, 19482, 1:2000).

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

Journal: bioRxiv

Article Title: miR-18a-5p upregulates Orai1 expression to promote vascular smooth muscle cell proliferation and neointimal hyperplasia after injury

doi: 10.64898/2026.01.20.700592

Figure Lengend Snippet: A. Representative images of immunofluorescence showing Orai1 (green), SARAF (red), and Dapi (blue) in a cross-section of the control and injured carotid arteries. Scale bar: 100 μm. B, C. Representative immunoblots (top) and summary data (bottom) showing the protein expression density of Orai1 (B) , and SARAF (C) normalized to its corresponding α-SMA. D. Representative images of Proximity ligation assay performed on a cross-section of control and injured arteries at 1 week post-surgery. Sections were incubated with mouse anti-Orai1 and rabbit anti-SARAF antibodies. Red fluorescent dots indicate proximity (< 40 nm) between Orai1 and SARAF. The nuclei were stained with DAPI (blue). Images were captured using a confocal microscope with 10X, 20X, and 63X objectives, with maximum z-projection of planes at 0.4 μm intervals. Scale bar: 100 μm. E. Co-immunoprecipitation of homogenized lysates from control and injured carotid arteries 1 week post-surgery with an anti-Orai1 antibody and analyzed by western blot using anti-STIM1 (Blot: STIM1), anti-SARAF (Blot: SARAF), and anti-Orai1 (Input: Orai1) antibodies as control. (n = pull of 5 arteries per conditions). 1W: 1 week; 2W: 2 weeks; 3W: 3 weeks post-injury. Data are presented as mean ± SD. (*), (**) and (****) indicate significance with p < 0.05, p < 0.01 and p < 0.0001, respectively.

Article Snippet: Afterwards, carotid sections were permeabilized and blocked with 10% goat serum and 3% bovine serum albumin in phosphate-buffered saline (PBS1X) with 0,1% Triton for 1 h. After blocking, the slides were incubated with primary antibodies: rabbit anti-CD31 (1:400, Novus Biologicals, US), mouse anti-αSMA (1:400, Sigma-Aldrich, San Luis, MI, US), rabbit anti-SARAF (1:200, Abcam, Cambridge, UK) or mouse anti-Orai1 (1:200, Novus Biologicals, Centennial, CO, US) at 4°C overnight.

Techniques: Immunofluorescence, Control, Western Blot, Expressing, Proximity Ligation Assay, Incubation, Staining, Microscopy, Immunoprecipitation

Journal: bioRxiv

Article Title: PIGBOS-CLCC1 Interaction Shapes Cellular Calcium Dynamics and Energy Metabolism

doi: 10.64898/2026.01.08.697870

Figure Lengend Snippet: (A) TG-induced ER Ca 2+ leak and SOCE recordings in HEK293T cells show reduced ER Ca 2+ leak and enhanced SOCE in PIGBOS-overexpressing cells ( n = 66) compared with mock controls ( n = 58). (B) PIGBOS knockdown ( n = 44) leaves ER Ca 2+ leak unchanged but markedly reduces SOCE relative to control cells ( n = 71). (C) Confocal images showing STIM1 and Orai1 puncta formation following TG-induced ER Ca 2+ store depletion in control and PIGBOS KO cells. (D and E) Quantification of STIM1 ( n = 7-18 cells) and Orai1 ( n = 6-18 cells) puncta before and after TG treatment. (F) Combined plasma membrane fluorescence intensity of STIM1 and Orai1 before and after TG stimulation. Each data point represents a manually defined ROI along the plasma membrane. Data represent mean ± SD. n.s., not significant; *P < 0.05, **P < 0.01, ***P < 0.001 (unpaired t test).

Article Snippet: Samples were resolved on 12% Tricine gels, transferred to PVDF membranes, blocked in 5% BSA, and probed with antibodies against PIGBOS (Abgenex #ABP121, 1:500), CLCC1 (Thermo Fisher #A305-668A-T, 1:1,000), STIM1 (CST #D88E10, 1:1,000), Orai1 (Novus #NBP1-85463, 1:1,000), VDAC (CST #4866, 1:1,000), and vinculin (CST #4650, 1:5,000), followed by HRP-conjugated secondary antibodies: anti-rabbit (CST #7074, 1:5,000) and anti-mouse (CST #7076, 1:5,000) and Clarity ECL detection (Bio-Rad #1705060).

Techniques: Knockdown, Control, Clinical Proteomics, Membrane, Fluorescence

Journal: bioRxiv

Article Title: PIGBOS-CLCC1 Interaction Shapes Cellular Calcium Dynamics and Energy Metabolism

doi: 10.64898/2026.01.08.697870

Figure Lengend Snippet: Figure 8. Model summarizing how PIGBOS coordinates ER-mitochondria-plasma membrane Ca 2+ signalling. PIGBOS engages CLCC1 via its C terminus to augment IP 3 R-dependent agonist-evoked ER Ca 2+ release, and STIM1-Orai1 coupling to potentiate SOCE, thereby amplifying cytosolic and mitochondrial Ca 2+ transients and supporting bioenergetic and redox homeostasis. Conversely, PIGBOS loss attenuates IP 3 R, STIM1, and Orai1 function, blunts ER-to-mitochondria Ca 2+ transfer, and promotes ROS accumulation, ATP depletion, and mitochondrial injury (figure created with BioRender).

Article Snippet: Samples were resolved on 12% Tricine gels, transferred to PVDF membranes, blocked in 5% BSA, and probed with antibodies against PIGBOS (Abgenex #ABP121, 1:500), CLCC1 (Thermo Fisher #A305-668A-T, 1:1,000), STIM1 (CST #D88E10, 1:1,000), Orai1 (Novus #NBP1-85463, 1:1,000), VDAC (CST #4866, 1:1,000), and vinculin (CST #4650, 1:5,000), followed by HRP-conjugated secondary antibodies: anti-rabbit (CST #7074, 1:5,000) and anti-mouse (CST #7076, 1:5,000) and Clarity ECL detection (Bio-Rad #1705060).

Techniques: Clinical Proteomics, Membrane