Journal: International Journal of Molecular Sciences

Article Title: STIM1 Deficiency Leads to Specific Down-Regulation of ITPR3 in SH-SY5Y Cells

doi: 10.3390/ijms21186598

Figure Lengend Snippet: Downregulation of ITPR3 in STIM1-deficient cells. ( a ) Total RNA was purified from undifferentiated wild-type (WT) and STIM1-KO cells, and the quantification of transcripts was evaluated from a Taqman Gene Expression Array (Ref. #4418932). Data from 3 different experiments are given in the (n = 3 for WT; n = 3 for KO). From these data, the threshold cycle (Ct) was calculated to evaluate the expression fold change as 2^(−ΔΔC t ) which is also plotted in the bar chart, as the fold-change of ITPR1/3 expression in STIM1-KO cells compared with wild-type cells. Expression of GAPDH was used as a housekeeping gene. ( b ) Quantification of ITPR1/2/3 transcripts was performed individually with specific primers (see Methods, ). The bar chart depicts the expression fold change of transcripts in STIM1-KO cells compared with wild-type cells. Data are mean ± S.D. from 2 different biological replicates with technical triplicates (n = 6). ( c – e ) ITPR3, ITPR1, and ITPR2 protein expression in whole cell lysates was quantified in wild-type and STIM1-KO cells. Beta-tubulin was used as a loading control. Data are mean ± S.D. from 3 independent experiments (for ITPR1) or 4 independent experiments (for ITPR2 and ITPR3). ( f ) ITPR3 protein expression (left panel) or total levels of ITPRs (right panel) were evaluated in osteosarcoma U2OS cells, as well as in wild-type and STIM1-KO SH-SY5Y cells by immunoblot. Beta-tubulin was used as a loading control. (***) Statistical significance p < 0.001.

Article Snippet: The rabbit polyclonal anti-STIM1 antibody (#4119) was from ProSci Inc. (Poway, CA, USA); the mouse monoclonal anti-red fluorescent protein (RFP, clone 6G6) was from Chromotek (Planegg-Martinsried, Germany); the mouse anti-beta tubulin antibody (clone TUB2.1) was from Sigma-Aldrich; the mouse anti-ITPR3 antibody (#610312), was from BD Biosciences (Franklin Lakes, NJ, USA); the mouse anti-ITPR2 antibody (sc-398434, clone A-5), the mouse anti-ITPR1/2/3 antibody (sc-377518, clone B-2), the mouse anti-ASCL4 (sc-365230, clone F-4), mouse anti-VDAC1 (sc-390996, clone B-6), and the mouse monoclonal anti-GAPDH antibodies were from Santa Cruz Biotechnology (Heidelberg, Germany); the rabbit anti-ITPR1 antibody (RBT-03) was a kind gift from Dr. Jan B. Parys (KU Leuven) who generated and characterized the antibody elsewhere [ ].

Techniques: Purification, Gene Expression, Expressing, Control, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: STIM1 Deficiency Leads to Specific Down-Regulation of ITPR3 in SH-SY5Y Cells

doi: 10.3390/ijms21186598

Figure Lengend Snippet: Downregulation of ITPR3 in differentiated STIM1-KO cells. ( a ) Quantification of ITPR1 , ITPR2 , and ITPR3 transcripts from differentiated STIM1-KO cells compared to wild-type cells. Data are mean ± S.D. from 2 different biological replicates with technical triplicates (n = 6). ( b ) ITPR3 protein expression in whole cell lysates was quantified in differentiated wild-type and STIM1-KO cells (beta-tubulin as a loading control). Data are mean ± S.D. from 2 independent experiments.

Article Snippet: The rabbit polyclonal anti-STIM1 antibody (#4119) was from ProSci Inc. (Poway, CA, USA); the mouse monoclonal anti-red fluorescent protein (RFP, clone 6G6) was from Chromotek (Planegg-Martinsried, Germany); the mouse anti-beta tubulin antibody (clone TUB2.1) was from Sigma-Aldrich; the mouse anti-ITPR3 antibody (#610312), was from BD Biosciences (Franklin Lakes, NJ, USA); the mouse anti-ITPR2 antibody (sc-398434, clone A-5), the mouse anti-ITPR1/2/3 antibody (sc-377518, clone B-2), the mouse anti-ASCL4 (sc-365230, clone F-4), mouse anti-VDAC1 (sc-390996, clone B-6), and the mouse monoclonal anti-GAPDH antibodies were from Santa Cruz Biotechnology (Heidelberg, Germany); the rabbit anti-ITPR1 antibody (RBT-03) was a kind gift from Dr. Jan B. Parys (KU Leuven) who generated and characterized the antibody elsewhere [ ].

Techniques: Expressing, Control

Journal: Poultry Science

Article Title: Goose astrovirus induces apoptosis and endoplasmic reticulum stress in gosling hepatocytes

doi: 10.1016/j.psj.2024.104600

Figure Lengend Snippet: qPCR Primer.

Article Snippet: The cells were incubated overnight at 4 °C with the primary antibody against IP3R (1:1000, Servicebio, Wuhan, China).

Techniques:

Journal: Poultry Science

Article Title: Goose astrovirus induces apoptosis and endoplasmic reticulum stress in gosling hepatocytes

doi: 10.1016/j.psj.2024.104600

Figure Lengend Snippet: GoAstV infection can cause liver ER calcium disorders. A. Changes of calcium content in liver tissues of two groups. B. The heatmap illustrates the expression levels of genes associated with ER calcium channels. C. Expression levels of mRNA for ER calcium channel-associated genes (IP3R, RYR3, CaMKII, and SERCA2α). D-F. Western blot analysis to quantify the relative expression levels of CaMKII and SERCA2 proteins. E-F. Immunohistochemical staining at 6 dpi and ImageJ analysis of positive expression of IP3R(Scale bar: 20 µm.B). All data are presented as M ± SE and evaluated using an independent samples t-test. P < 0.05 (*). P < 0.01 (**).

Article Snippet: The cells were incubated overnight at 4 °C with the primary antibody against IP3R (1:1000, Servicebio, Wuhan, China).

Techniques: Infection, Expressing, Western Blot, Immunohistochemical staining, Staining

Journal: Bioactive Materials

Article Title: Amorphous calcium zinc phosphate promotes macrophage-driven alveolar bone regeneration via modulation of energy metabolism and mitochondrial homeostasis

doi: 10.1016/j.bioactmat.2025.06.053

Figure Lengend Snippet: ACZP reverses the pro-inflammatory polarization phenotype of macrophages induced by HA through inhibiting the IP3R/MCU axis: (A) GO bubble plots showed the calcium ion transport-related biological function differences of macrophages between the HA and Control groups. Dot size and color correspond to the enrichment P -value of that term (after -log10 transformation). (B) Heatmap of the expression of calcium ion transport-related genes in all macrophage clusters between the HA and Control groups. (C) Violin plot of the expression of the Micu1 gene in macrophages of the HA and Control groups. (D) UMAP atlas showed the expression of Micu1 in macrophages of the HA and Control groups. (E) Two-dimensional image of IP3R and MCU molecular with or without HA or ACZP docking simulation. (F) Representative images of the co-localization of the mitochondria and ER in macrophages stained with the Mito-Tracker and ER-Tracker (green: mitochondria; red: ER; scale bar: 10 μm; 2 μm (inset); The arrow pointed to the MAMs structure.). (G) Immuno-electron microscopy showed the expression of IP3R and MCU antibodies between the mitochondria and ER. (scale bar: 500 nm). (H) The morphology of the mitochondria and ER of Control, HA and ACZP groups were analyzed by TEM, and the representative images were displayed. (green: mitochondria; purple: ER; scale bar: 2 μm (left); 500 nm (right)). (I–J) The distance between the ER and mitochondria (n = 30) and the mitochondrial perimeter (n = 30) were quantified and analyzed, respectively. The data were shown as the mean ± SD; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001 indicated significant differences between the indicated columns (one-way ANOVA). (K) Schematic diagram of the calcium ion transport channel between the mitochondria and ER in macrophages.

Article Snippet: Sections were incubated overnight with specific antibodies (CD68, MCU, IP3R; Scanco Medical AG, ZH, CH) at 4 °C.

Techniques: Control, Transformation Assay, Expressing, Staining, Immuno-Electron Microscopy

Journal: Bioactive Materials

Article Title: Amorphous calcium zinc phosphate promotes macrophage-driven alveolar bone regeneration via modulation of energy metabolism and mitochondrial homeostasis

doi: 10.1016/j.bioactmat.2025.06.053

Figure Lengend Snippet: ACZP promotes the repair of alveolar bone defects in New Zealand white rabbits: (A) Schematic diagram of nanomaterials for treating rabbit alveolar bone defects. (B) Representative Micro-CT, H&E, and Masson staining of rabbit alveolar bone 4 weeks after implantation in the Control group, HA group and ACZP group (scale bar: 1 mm (Micro-CT); 500 μm (H&E and Masson staining)). (C) Representative Micro-CT, H&E, and Masson staining of rabbit alveolar bone 12 weeks after implantation in the Control group, HA group and ACZP group (scale bar: 1 mm (Micro-CT); 500 μm (H&E and Masson staining)). (D–E) Semi-quantification of BV/TV, Tb.N, Tb.Th and Tb.Sp in different groups at 4 (left) and 12 (right) weeks. ( n = 3). The data were shown as the mean ± SD ( n = 3); ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001 indicated significant differences between the indicated columns (one-way ANOVA). (F) Representative immunofluorescence image of MCU staining (yellow) and IP3R staining (green) in alveolar bone defect after 4 weeks of implantation. (scale bar: 200 μm).

Article Snippet: Sections were incubated overnight with specific antibodies (CD68, MCU, IP3R; Scanco Medical AG, ZH, CH) at 4 °C.

Techniques: Micro-CT, Staining, Control, Immunofluorescence