Journal: bioRxiv

Article Title: FAM84B facilitates tau propagation via RYR3-mediated exocytosis in response to neuroinflammation

doi: 10.64898/2025.12.28.696549

Figure Lengend Snippet: (A) Differentially expressed genes in HEK293 cells in response to FAM84B KO (n=3 each). Genes with p < 0.05 and >2-fold expression change compared to control are marked with red dots. (B) Differentially expressed genes in HEK293 cells in response to FAM84B OE (n=3 each). Genes with p < 0.05 and >2-fold expression change compared to control are marked with red dots. (C) Differentially expressed genes in tau-expressing SH-SY5Y cells in response to FAM84B OE (n=3 each). Genes with p < 0.05 and >1.7-fold expression change compared to cells expressing tau alone are marked with red dots. (D) Venn diagram showing the number of shared genes between RNA-seq datasets in (A–C), with overlapping genes represented in distinct colors (red, green, black, blue). Only the names of membrane-related genes among the shared genes are noted. (E) GO analysis showing cellular compartments of genes identified in (D). The GO terms based on the number of genes are shown. (F) Levels of RYR3 and actin in iPSC-derived CN from HC and patients with AD (n=3 each). (G) Levels of RYR3 and actin in SH-SY5Y cells expressing tau alone or tau with FAM84B-Myc (n=3 each). (H) Colocalization (arrow) of FAM84B and calcium in SH-SY5Y cells expressing tau with mCherry control (C1), and in cells expressing tau with mCherry-FAM84B treated with either DMSO or dantrolene (n=4 each). (I) PHF-1 spread from SH-SY5Y cells expressing Tau-p2a-eGFP with FAM84B to neighboring cells in response to treatment with either DMSO or dantrolene (n=4 each). Arrows indicate recipient cells (red) to which tau has been transmitted. Solid boxes in the images indicate magnified areas. Results are presented as mean ± SEM. Co-localization analysis. (H) Pearson correlation coefficient. Statistical analyses: (A-C, F-I) unpaired two-tailed t -test with Welch’s correction; ns: not significant, *p < 0.05, **p < 0.01, ***p < 0.001. Abbreviations: KO, knockout; OE, overexpression; GO, gene ontology; CN, cortical neuron; HC, healthy control; AD, Alzheimer’s disease; PHF-1, phosphorylated tau at residues S396/S404; FOV, field of view.

Article Snippet: Following blocking, membranes were incubated overnight at 4°C with primary antibodies (1:1,000 dilution), including: PHF-1 (Peter Davies Lab), Tau5 (Gail VW Johnson Lab), Tau (Dako, Agilent Technologies, Inc., 0024), Myc-tag (Cell Signaling Technology, 2276), FAM84B (OriGene, TA501992; Proteintech Group, Inc., Rosemont, IL, USA, 18421-1-AP), Alix (Cell Signaling Technology, 2171), Caveolin-1 (Cell Signaling Technology, 3267), RYR3 (Alomone Labs, Jerusalem, Israel, ARR003), Clathrin-heavy chain (CHC, Cell Signaling Technology, 4796), EEA1 (Cell Signaling Technology, 3288), Rab5 (Cell Signaling Technology, 3547), Rab7 (Cell Signaling Technology, 9367), Rab9 (Cell Signaling Technology, 5118), Rab11 (Cell Signaling Technology, 5589), pSTAT3 (Y705) (Cell Signaling Technology, 9145), STAT3 (Cell Signaling Technology, 9139), p62 (Cell Signaling Technology, 8025), LC3 (Cell Signaling Technology, 3868), HSP70 (Cell Signaling Technology, 4876), LAMP-1 (BD Biosciences, Franklin Lakes, NJ, USA, 611042), LAMP-2 (Sigma-Aldrich, L0668), and beta-actin (Merck Millipore, MAB1501).

Techniques: Expressing, Control, RNA Sequencing, Membrane, Derivative Assay, Two Tailed Test, Knock-Out, Over Expression

Journal: bioRxiv

Article Title: FAM84B facilitates tau propagation via RYR3-mediated exocytosis in response to neuroinflammation

doi: 10.64898/2025.12.28.696549

Figure Lengend Snippet: (A) Venn diagram showing the seven shared transcriptional cofactors between the ENCODE ChIP-seq and CHEA ChIP-seq datasets. The genes are categorized into functional terms and noted. (B) Genomic information of the FAM84B gene obtained from the UCSC Genome Browser. Transcription cofactor clusters identified from the ENCODE ChIP-seq data, with those likely to bind to the regulatory elements of FAM84B, as identified by FANTOM5, highlighted. (C) Levels of phosphorylated STAT3 (Y705) in the cerebral cortex of HC (n=3) and patients with VD (n=3), CBD (n=3), PSP (n=4), PART (n=3), and AD (n=6). (D) Levels of phosphorylated STAT3 (Y705) and total STAT3 in iPSC-derived CN from HC and patients with AD (n=3 each). (E) Levels of phosphorylated STAT3 (Y705), total STAT3, FAM84B, and actin in SH-SY5Y cells treated with either control vehicle or IL-6 (n=3 each). (F) Changes in the levels of phosphorylated STAT3 (Y705), total STAT3, FAM84B, and actin in SH-SY5Y cells expressing STAT3-WT or STAT3-DN in response to treatment with either control vehicle or IL-6 (n=3 each). (G) PHF-1 spread from Tau-p2a-eGFP expressing SH-SY5Y cells to neighboring cells in response to treatment with either control vector or IL-6 (n=4 each). Arrows indicate recipient cells (red) to which tau has been transmitted. Solid boxes in the images indicate magnified areas. (H) PHF-1 spread from IL-6 treated SH-SY5Y cells expressing Tau-p2a-eGFP to neighboring cells in response to treatment with either DMSO or dantrolene (n=4 each). Arrows indicate recipient cells (red) to which tau has been transmitted. Solid boxes in the images indicate magnified areas. (I) Changes in the levels of PHF-1, total tau, phosphorylated STAT3 (Y705), total STAT3, FAM84B (FAM84B-Myc), RYR3, and actin in iPSC-derived CN from HC after infection with AAV and subsequent treatments (n=3 each). (J) Changes in the levels of phosphorylated tau (T181 and S396) in the CM of iPSC-derived CN from HC after infection with AAV and subsequent treatments (n=8 each). In (I–J), the experimental groups are: (1) CN infected with AAV containing tau, (2) CN infected with AAV containing tau and FAM84B-Myc, (3) CN infected with AAV containing tau and treated with IL-6 and TNF-α, (4) CN infected with AAV containing tau and FAM84B-Myc and treated with dantrolene, and (5) CN infected with AAV containing tau, treated with IL-6 and TNF-α, followed by treatment with dantrolene. Results are presented as mean ± SEM. Statistical analyses: (C) ordinary two-way ANOVA; (D-J) unpaired two-tailed t -test with Welch’s correction; ns: not significant, *p < 0.05, **p < 0.01, and ***p < 0.001. Abbreviations: HC, healthy control; VD, vascular dementia; CBD, corticobasal degeneration; PSP, progressive supranuclear palsy; PART, primary age-related tauopathy; AD, Alzheimer’s disease; CN, cortical neuron; WT, wild type; DN, dominant negative; PHF-1, phosphorylated tau at S396/S404 residue; FOV, field of view; AAV, adeno-associated virus; CM, conditioned media.

Article Snippet: Following blocking, membranes were incubated overnight at 4°C with primary antibodies (1:1,000 dilution), including: PHF-1 (Peter Davies Lab), Tau5 (Gail VW Johnson Lab), Tau (Dako, Agilent Technologies, Inc., 0024), Myc-tag (Cell Signaling Technology, 2276), FAM84B (OriGene, TA501992; Proteintech Group, Inc., Rosemont, IL, USA, 18421-1-AP), Alix (Cell Signaling Technology, 2171), Caveolin-1 (Cell Signaling Technology, 3267), RYR3 (Alomone Labs, Jerusalem, Israel, ARR003), Clathrin-heavy chain (CHC, Cell Signaling Technology, 4796), EEA1 (Cell Signaling Technology, 3288), Rab5 (Cell Signaling Technology, 3547), Rab7 (Cell Signaling Technology, 9367), Rab9 (Cell Signaling Technology, 5118), Rab11 (Cell Signaling Technology, 5589), pSTAT3 (Y705) (Cell Signaling Technology, 9145), STAT3 (Cell Signaling Technology, 9139), p62 (Cell Signaling Technology, 8025), LC3 (Cell Signaling Technology, 3868), HSP70 (Cell Signaling Technology, 4876), LAMP-1 (BD Biosciences, Franklin Lakes, NJ, USA, 611042), LAMP-2 (Sigma-Aldrich, L0668), and beta-actin (Merck Millipore, MAB1501).

Techniques: ChIP-sequencing, Functional Assay, Derivative Assay, Control, Expressing, Plasmid Preparation, Infection, Two Tailed Test, Dominant Negative Mutation, Residue, Virus

Journal: American Journal of Physiology - Cell Physiology

Article Title: Purinergic agonists increase [Ca 2+ ] i in rat conjunctival goblet cells through ryanodine receptor type 3

doi: 10.1152/ajpcell.00291.2024

Figure Lengend Snippet: Effect of blocking L-type calcium channels with nifedipine and ryanodine receptors with dantrolene on ATP and BzATP-induced peak [Ca 2+ ] i . Cultured rat conjunctival goblet cells were preincubated with nifedipine (Nif) (10 −5 M) for 15 min and/or dantrolene (Da) (10 −5 M) for 30 min before stimulation with ATP (10 −5 M) ( A ) or BzATP (10 −4 M) ( B ). A and B : the average [Ca 2+ ] i over time. Arrows represent the addition of agonists. C : the peak increase of [Ca 2+ ] i above baseline. ATP: bars to the left of the central vertical dotted line in C , BzATP: bars to the right of the central vertical dotted line in C . Data are presented as means ± SE. n = 3. BzATP, benzoylbenzoyl-ATP; [Ca 2+ ] i , intracellular calcium concentration. *Statistically significant difference from agonist alone.

Article Snippet: Cells were incubated with monoclonal mouse anti-RyR1/2 antibody (Thermo Fisher Scientific, Cat. No. MA3-916, RRID:AB_2183054) or rabbit anti-RyR3 antibody (Alomone Labs, Cat. No. ARR-003, RRID:AB_2040186) at a 1:50 or 1:100 dilution overnight at 4°C.

Techniques: Blocking Assay, Cell Culture, Concentration Assay

Journal: American Journal of Physiology - Cell Physiology

Article Title: Purinergic agonists increase [Ca 2+ ] i in rat conjunctival goblet cells through ryanodine receptor type 3

doi: 10.1152/ajpcell.00291.2024

Figure Lengend Snippet: RT-qPCR of the ryanodine receptor 3 (RyR3) subtype in rat and human conjunctival goblet cells (CGCs). Total RNA was isolated from rat and human CGCs and reverse-transcribed. cDNA amplification confirmed the presence of RyR3 mRNA in rat ( A ) but not in human ( B ) CGCs. Relative amount of RyR3 transcript compared with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was measured by quantitative PCR and is shown on y -axis. n = 3. RT-qPCR, quantitative reverse transcription polymerase chain reaction. *Statistically significant difference from 0.

Article Snippet: Cells were incubated with monoclonal mouse anti-RyR1/2 antibody (Thermo Fisher Scientific, Cat. No. MA3-916, RRID:AB_2183054) or rabbit anti-RyR3 antibody (Alomone Labs, Cat. No. ARR-003, RRID:AB_2040186) at a 1:50 or 1:100 dilution overnight at 4°C.

Techniques: Quantitative RT-PCR, Isolation, Reverse Transcription, Amplification, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction

Journal: American Journal of Physiology - Cell Physiology

Article Title: Purinergic agonists increase [Ca 2+ ] i in rat conjunctival goblet cells through ryanodine receptor type 3

doi: 10.1152/ajpcell.00291.2024

Figure Lengend Snippet: Proposed regulation of intracellular calcium levels by ryanodine receptor 3 (RyR3) in cultured rat conjunctival goblet cells. ATP and BzATP may increase [Ca 2+ ] i : 1 ) directly by activating RyR3, 2 ) by activating purinergic 1 (P1) receptors after the enzymatic conversion to metabolites, 3 ) by the activation of purinergic 2 (P2) receptors directly, and 4 ) by activating L-type calcium channels through membrane depolarization. These mechanisms can then either directly increase [Ca 2+ ] i through the influx of extracellular Ca 2+ and/or through the activation of RyR3 on an intracellular compartment such as the ER to release Ca 2+ . BzATP, benzoylbenzoyl-ATP; [Ca 2+ ] i , intracellular calcium concentration; CGCs, conjunctival goblet cells; CICR, calcium-induced calcium release.

Article Snippet: Cells were incubated with monoclonal mouse anti-RyR1/2 antibody (Thermo Fisher Scientific, Cat. No. MA3-916, RRID:AB_2183054) or rabbit anti-RyR3 antibody (Alomone Labs, Cat. No. ARR-003, RRID:AB_2040186) at a 1:50 or 1:100 dilution overnight at 4°C.

Techniques: Cell Culture, Activation Assay, Membrane, Concentration Assay

Journal: The Journal of Poultry Science

Article Title: Egg Development After In Vitro Insemination in Japanese Quail ( Coturnix japonica )

doi: 10.2141/jpsa.2023001

Figure Lengend Snippet: Presence of CCNB1, c-MOS, ITPR1, and RYR3 proteins in quail eggs and their down-regulated expression after ICSI. (A) Detection of CCNB1, c-MOS, ITPR1, and RYR3 proteins in ovulated quail eggs. (B) Western blot analysis 3 h after ICSI with injection of PLCZ1, CS, and ACO2 cRNA or the solvent alone. (C) Western blot analysis 3 h after ICSI with PLCZ1 cRNA alone, CS and ACO2 cRNAs, or the solvent alone. (D) Quantification of immunoreactivity after ICSI. Band intensities were quantified and expressed as the means ± standard deviations of three samples. Values with different letters are significantly different (P < 0.01).

Article Snippet: Rabbit anti-rat ITPR1 antibody (Alomone Labs Ltd., Jerusalem, Israel) and mouse anti-chicken RYR antibody (GeneTex, Inc., Irvine, CA, USA) were used to detect ITPR1 and RYR3, respectively.

Techniques: Expressing, Western Blot, Injection, Solvent

Journal: The Journal of Poultry Science

Article Title: Egg Development After In Vitro Insemination in Japanese Quail ( Coturnix japonica )

doi: 10.2141/jpsa.2023001

Figure Lengend Snippet: Changes in ITPR1, RYR3, CCNB1, and c-MOS protein expression levels in quail eggs following in vitro insemination with different sperm numbers. (A–C) Western blot analysis 3 h after in vitro insemination with 2 × 10 2 (A), 2 × 10 3 (B), or 2 × 10 4 (C) sperm, respectively. Numbers 1–6 represent each of the six eggs treated with sperm. Egg extracts after ICSI or PBS treatment were used as the control. (D) Quantification of immunoreactivity after in vitro insemination. Band intensities were quantified and are expressed as means ± standard deviations. The sample number of PBS- and ICSI-treated eggs is three each. Band intensities in these three samples were measured in each membrane.

Article Snippet: Rabbit anti-rat ITPR1 antibody (Alomone Labs Ltd., Jerusalem, Israel) and mouse anti-chicken RYR antibody (GeneTex, Inc., Irvine, CA, USA) were used to detect ITPR1 and RYR3, respectively.

Techniques: Expressing, In Vitro, Western Blot, Control, Membrane

Journal: International Journal of Molecular Sciences

Article Title: Transplantation of Neural Precursors Derived from Induced Pluripotent Cells Preserve Perineuronal Nets and Stimulate Neural Plasticity in ALS Rats

doi: 10.3390/ijms21249593

Figure Lengend Snippet: List of primary antibodies.

Article Snippet: Anti-Ryanodine Receptor 3 , Ryanodine Receptor 3/ICC , Rabbit polyclonal , 1:100 , Alomone Labs.

Techniques: Marker