ta326547  (OriGene)

Journal: The Journal of Headache and Pain

Article Title: ER stress-induced ITPR1/ANO1 signaling drives trigeminal neuropathic pain through calcium-dependent neuronal hyperexcitability

doi: 10.1186/s10194-025-02231-9

Figure Lengend Snippet: ITPR1 is increased in the TG after pIONT and mediates Ca 2+ release. ( A ) Time course of Itpr1 mRNA expression in TG of naive, sham-operated, and pIONT-operated mice. *** P < 0.001. Student’s t-test, n = 7 or 8 mice/group. ( B ) Representative Western blot of ITPR1 protein expression in TG 10 days post-pIONT. ( C ) Quantification of ITPR1 protein levels showing significant upregulation following pIONT. * P < 0.05. Student’s t-test, n = 4 mice/group ( D ) Representative images of ITPR1 staining in the TG of naive, sham-, and pIONT-operated mice. Double staining of ITPR1 with TUJ1, IBA-1, FABP7, CGRP, NF200, and IB4. ( E ) Quantitative analysis of ITPR1 fluorescence intensity. * P < 0.05, One-way ANOVA followed by the Bonferroni test, 3 mice/group. ( F ) Percentage of ITPR1 co-localization with cellular markers (TUJ1, FABP7, and IBA-1). ( G ) Percentage of ITPR1 co-localization with different neuron type markers (CGRP, NF200, and IB4). ( H ) Example traces of neuronal Ca²⁺ dynamics during ATP bath application, comparing NC siRNA- and Itpr1 siRNA-treated pIONT groups. ( I ) Representative traces of ATP-induced Ca²⁺ signals (ΔF/F₀) in NC siRNA- and Itpr1 siRNA-treated TG neurons. ( J ) Quantification showing Itpr1 siRNA significantly attenuates ATP-induced Ca²⁺ signaling. NC: n = 277 cells; Itpr1 siRNA: n = 200 cells; *** P < 0.001. Student’s t-test. 3 mice/group

Article Snippet: The ITPR1 agonist Adenophostin A (MCE; HY-142117), ITPR1 inhibitor 2-APB (Sigma‒Aldrich; Cat. No. CAS524958), ER stress inhibitor 4-PBA (Sigma‒Aldrich; CAS1821121), and ANO1 blocker MONNA (Sigma‒Aldrich; SML0902) were dissolved in 1% DMSO.

Techniques: Expressing, Western Blot, Staining, Double Staining, Fluorescence

Journal: The Journal of Headache and Pain

Article Title: ER stress-induced ITPR1/ANO1 signaling drives trigeminal neuropathic pain through calcium-dependent neuronal hyperexcitability

doi: 10.1186/s10194-025-02231-9

Figure Lengend Snippet: ITPR1 expression is regulated by the transcription factor Runx2. ( A ) Heatmap of transcription factor expression profiles in TG following pIONT. ( B ) JASPAR database prediction of RUNX2 binding sites in the Itpr1 promoter region. ( C ) Dual-luciferase reporter assay showing RUNX2 inhibition suppresses Itpr1 promoter activity. *** P < 0.001. One-way ANOVA followed by the Bonferroni test. ( D - E ) qPCR analysis of Runx2 ( D ) and Itpr1 ( E ) mRNA levels after TM injection. * P < 0.05. Student’s t-test. ( F ) Runx2 mRNA expression in TG post-pIONT. ** P < 0.01. Student’s t-test, n = 6 mice/group. ( G ) Representative Western blot of RUNX2 protein in TG 10 days post-pIONT. ( H ) Quantification confirms pIONT-induced RUNX2 upregulation. ** P < 0.01. Student’s t-test, n = 5 mice/group. ( I - J ) Mechanical allodynia (0.02 g, 0.16 g stimuli) showing that single-dose Runx2 siRNA (3 µg, day 0) attenuates pIONT-induced hypersensitivity. * P < 0.05. Two-way ANOVA followed by the Bonferroni test, n = 7 mice/group. ( K - L ) Runx2 ( K ) and Itpr1 ( L ) mRNA levels post-pIONT with Runx2 siRNA. * P < 0.05, ** P < 0.01. Student’s t-test, n = 6 mice/group. ( M , N ) A single dose injection of Runx2 siRNA (3 µg, day 7) attenuated pIONT-induced mechanical allodynia. * P < 0.05, ** P < 0.01. Two-way ANOVA followed by the Bonferroni test, n = 7 mice/group. ( O - P ) Runx2 ( O ) and Itpr1 ( P ) mRNA after late Runx2 siRNA intervention. * P < 0.05. Student’s t-test, n = 6 mice/group

Article Snippet: The ITPR1 agonist Adenophostin A (MCE; HY-142117), ITPR1 inhibitor 2-APB (Sigma‒Aldrich; Cat. No. CAS524958), ER stress inhibitor 4-PBA (Sigma‒Aldrich; CAS1821121), and ANO1 blocker MONNA (Sigma‒Aldrich; SML0902) were dissolved in 1% DMSO.

Techniques: Expressing, Binding Assay, Luciferase, Reporter Assay, Inhibition, Activity Assay, Injection, Western Blot

Journal: The Journal of Headache and Pain

Article Title: ER stress-induced ITPR1/ANO1 signaling drives trigeminal neuropathic pain through calcium-dependent neuronal hyperexcitability

doi: 10.1186/s10194-025-02231-9

Figure Lengend Snippet: ITPR1 mediates ERK activation, inflammatory mediator expression, and neuronal hyperexcitability in the TG after pIONT . ( A , B ) Western blot analysis demonstrating pIONT-induced pERK upregulation is attenuated by Itpr1 siRNA (A) or 2-APB (B) treatment. * P < 0.05, ** P < 0.01, Student’s t-test, n = 3 mice/group. ( C ) qPCR analysis showing Itpr1 siRNA suppresses pIONT-induced upregulation of inflammatory cytokines in TG. * P < 0.05, ** P < 0.01, *** P < 0.001. Student’s t-test, n = 6 mice/group. ( D ) qPCR analysis revealing 2-APB-mediated reduction of pIONT-induced cytokine expression in TG. * P < 0.05, ** P < 0.01. Student’s t-test, n = 6 mice/group. ( E ) Representative action potential (AP) traces evoked by rheobase current in TG neurons from Sham, pIONT, pIONT-NC siRNA, and pIONT- Itpr1 siRNA at 10 days after surgery. ( F ) Itpr1 siRNA prevents pIONT-induced reduction in AP rheobase. * P < 0.05, ** P < 0.01, Student’s t-test. n = 8–9 neurons/group from 3–4 mice. ( G ) Representative traces of APs evoked by depolarizing current steps across treatment groups. ( H ) Itpr1 siRNA significantly reduces pIONT-induced increase in AP firing. *** P < 0.001. Two-way ANOVA followed by the Bonferroni test. n = 8–9 neurons/group from 3–4 mice. ( I-J ) pIONT decreases resting membrane potential and threshold potential, which are unaffected by Itpr1 siRNA. ** P < 0.01, *** P < 0.001. Student’s t-test. n = 8–9 neurons/group from 3–4 mice

Article Snippet: The ITPR1 agonist Adenophostin A (MCE; HY-142117), ITPR1 inhibitor 2-APB (Sigma‒Aldrich; Cat. No. CAS524958), ER stress inhibitor 4-PBA (Sigma‒Aldrich; CAS1821121), and ANO1 blocker MONNA (Sigma‒Aldrich; SML0902) were dissolved in 1% DMSO.

Techniques: Activation Assay, Expressing, Western Blot, Membrane

Journal: The Journal of Headache and Pain

Article Title: ER stress-induced ITPR1/ANO1 signaling drives trigeminal neuropathic pain through calcium-dependent neuronal hyperexcitability

doi: 10.1186/s10194-025-02231-9

Figure Lengend Snippet: ITPR1 contributes to pIONT–induced mechanical allodynia and chewing disorder . ( A , B ) Mechanical allodynia assessment (0.02 g, 0.16 g stimuli) showing single-dose Itpr1 siRNA (3 µg, day 0) attenuates pIONT-induced hypersensitivity. * P < 0.05, ** P < 0.01. Two-way ANOVA followed by the Bonferroni test, n = 6 mice/group. ( C , D ) Delayed intervention with Itpr1 siRNA (3 µg, day 7 post-pIONT) similarly reverses mechanical allodynia. * P < 0.05, ** P < 0.01. Two-way ANOVA followed by the Bonferroni test, n = 6 mice/group. ( E , F ) Early intra-TG 2-APB administration (2 µg, day 0) ameliorates pIONT-induced mechanical hypersensitivity. * P < 0.05. Two-way ANOVA followed by the Bonferroni test, n = 8 mice/group. ( G - H )Therapeutic 2-APB treatment (2 µg, day 7) maintains analgesic efficacy against established allodynia. * P < 0.05, ** P < 0.01. Two-way ANOVA followed by the Bonferroni test, n = 8 mice/group. ( I ) Representative pine wood blocks showing gnawing patterns in sham vs. pIONT groups. ( J ) Quantification of the weight reduction in sham-operated and pIONT-operated mice. *** P < 0.001. Two-way ANOVA followed by the Bonferroni test, n = 6 mice/group. ( K ) Wood block specimens from NC siRNA vs. Itpr1 siRNA-treated pIONT mice. ( L ) Quantification of the weight reduction in Itpr1 siRNA- or NC siRNA-treated mice. ** P < 0.01. Two-way ANOVA followed by the Bonferroni test, n = 6 mice/group

Article Snippet: The ITPR1 agonist Adenophostin A (MCE; HY-142117), ITPR1 inhibitor 2-APB (Sigma‒Aldrich; Cat. No. CAS524958), ER stress inhibitor 4-PBA (Sigma‒Aldrich; CAS1821121), and ANO1 blocker MONNA (Sigma‒Aldrich; SML0902) were dissolved in 1% DMSO.

Techniques: Blocking Assay

Journal: The Journal of Headache and Pain

Article Title: ER stress-induced ITPR1/ANO1 signaling drives trigeminal neuropathic pain through calcium-dependent neuronal hyperexcitability

doi: 10.1186/s10194-025-02231-9

Figure Lengend Snippet: ITPR1 couples with the ANO1 channel . ( A ) Co-immunoprecipitation analysis of ITPR1-ANO1 complexes in TG lysates. Top: ITPR1 pulldown using ANO1 antibody; Bottom: ANO1 pulldown using ITPR1 antibody. Controls: Input (positive), IgG (negative). ( B ) Representative Western blot of ANO1 protein expression in TG 10 days post-pIONT. ( C ) Quantification reveals significant ANO1 upregulation in the pIONT group. * P < 0.05. Student’s t-test. n = 4 mice/group. ( D ) Double staining of ITPR1 with ANO1 in TG 10 days post-pIONT. ( E ) Bar charts showed the percentage of double-stained TG neurons among total numbers of ITPR1 + labeled neurons. n = 4 mice. ( F ) Representative AdA-induced inward currents in TG neurons from sham, pIONT, pIONT + veh, and pIONT + MONNA group mice. ( G ) Current amplitude quantification. ** P < 0.01, *** P < 0.001, Student’s t-test. n = 13 neurons/group from 3–4 mice. ( H - I ) A single dose of AdA in TG of naïve mice induced mechanical allodynia. ( J-K ) Mechanical allodynia after repeated injections of AdA or AdA + MONNA in TG of naive mice. * P < 0.05, ** P < 0.01, *** P < 0.001. Two-way ANOVA followed by the Bonferroni test. n = 7 mice/group

Article Snippet: The ITPR1 agonist Adenophostin A (MCE; HY-142117), ITPR1 inhibitor 2-APB (Sigma‒Aldrich; Cat. No. CAS524958), ER stress inhibitor 4-PBA (Sigma‒Aldrich; CAS1821121), and ANO1 blocker MONNA (Sigma‒Aldrich; SML0902) were dissolved in 1% DMSO.

Techniques: Immunoprecipitation, Western Blot, Expressing, Double Staining, Staining, Labeling

Journal: The Journal of Headache and Pain

Article Title: ER stress-induced ITPR1/ANO1 signaling drives trigeminal neuropathic pain through calcium-dependent neuronal hyperexcitability

doi: 10.1186/s10194-025-02231-9

Figure Lengend Snippet: ITPR1 couples with ANO1 to regulate neuronal excitability and TNP pathogenesis. ( A ) Representative traces of APs evoked by rheobase current in TG neurons from NC siRNA- or Ano1 siRNA-treated mice (10 days post-surgery). ( B ) Ano1 siRNA significantly increased AP rheobase vs. NC siRNA. * P < 0.05. Student’s t-test. n = 9–10 neurons/group from 3–4 mice. ( C ) Resting membrane potential depolarization following Ano1 siRNA. ** P < 0.01. Student’s t-test. n = 9–10 neurons/group from 3–4 mice. ( D ) Representative AP trains evoked by depolarizing current steps. ( E ) Ano1 siRNA reduced the number of APs. *** P < 0.001. Two-way ANOVA followed by the post hoc Bonferroni test. n = 9–10 neurons/group from 3–4 mice. ( F ) The thresholds were not affected by Ano1 siRNA treatment. ( G , H ) A single dose of Ano1 siRNA (3 µg) injection on day 0 post-pIONT significantly attenuated mechanical allodynia. ( I , J ) A single dose of Ano1 siRNA (3 µg) injection on day 7 post-pIONT significantly attenuated mechanical allodynia. ( K, L ) Intra-TG injection of MONNA (2 µg) on pIONT day 0 attenuated mechanical allodynia. ( M, N ) Intra-TG injection of MONNA (2 µg) on pIONT day 7 attenuated mechanical allodynia. ** P < 0.01, *** P < 0.001. Two-way ANOVA followed by the post hoc Bonferroni test. n = 7 mice/group

Article Snippet: The ITPR1 agonist Adenophostin A (MCE; HY-142117), ITPR1 inhibitor 2-APB (Sigma‒Aldrich; Cat. No. CAS524958), ER stress inhibitor 4-PBA (Sigma‒Aldrich; CAS1821121), and ANO1 blocker MONNA (Sigma‒Aldrich; SML0902) were dissolved in 1% DMSO.

Techniques: Membrane, Injection

Journal: The Journal of Headache and Pain

Article Title: ER stress-induced ITPR1/ANO1 signaling drives trigeminal neuropathic pain through calcium-dependent neuronal hyperexcitability

doi: 10.1186/s10194-025-02231-9

Figure Lengend Snippet: Schematic summarizing the mechanism by which ITPR1 promotes trigeminal neuropathic pain . Following trigeminal nerve injury, ER stress is induced in TG neurons, which upregulates ITPR1 expression via the transcription factor RUNX2, leading to enhanced Ca²⁺ release from ER stores. These localized Ca²⁺ signals activate the ERK signaling pathway, resulting in hyperexcitability of sensory neurons and increased expression of pro-inflammatory mediators, including TNF-α, IL-1β, IL-6, and CCL2. Additionally, ITPR1 functionally couples with ANO1, where ITPR1-mediated Ca²⁺ release potentiates ANO1 activation. This interaction promotes Cl⁻ efflux and subsequent membrane depolarization, further lowering the activation threshold of sensory neurons and perpetuating trigeminal neuropathic pain. The illustration was generated using BioRender

Article Snippet: The ITPR1 agonist Adenophostin A (MCE; HY-142117), ITPR1 inhibitor 2-APB (Sigma‒Aldrich; Cat. No. CAS524958), ER stress inhibitor 4-PBA (Sigma‒Aldrich; CAS1821121), and ANO1 blocker MONNA (Sigma‒Aldrich; SML0902) were dissolved in 1% DMSO.

Techniques: Expressing, Activation Assay, Membrane, Generated

Journal: International Journal of Molecular Sciences

Article Title: Perinatal Ethanol Exposure Induces Astrogliosis and Decreases GRP55/PEA-Mediated Neuroprotection in Hippocampal Astrocytes of the 3×Tg Alzheimer’s Animal Model

doi: 10.3390/ijms262211154

Figure Lengend Snippet: mRNA expression of the Ca 2+ signaling factors P2rx5 ( A ), Mcu ( B ), Nsmf ( C ), Itpr1 ( D ), Gls ( E ), and Gls2 ( F ) in the hippocampal astrocytes from 3×Tg-AD offspring of both sexes born to control and PEE mothers. Data are expressed as the mean ± S.E.M. ( n = 3–6). Relevant p values from the two-way ANOVA are shown below each graph, with significant p -values highlighted in bold. Tukey or single-effect analysis: ** p < 0.01. See also effect sizes in the .

Article Snippet: Itpr1 , Mm00439907 , NM_010585.5 , 58.

Techniques: Expressing, Control

Journal: Molecular Neurobiology

Article Title: Metformin Ameliorates Early Brain Injury After Subarachnoid Hemorrhage Via Improving Endoplasmic Reticulum Stress and Mitochondrial Stress-Mediated Ca 2+ Imbalance

doi: 10.1007/s12035-025-05558-1

Figure Lengend Snippet: ERSand MS mediated the IP3R1–GRP75–VDAC1 Ca2 + channeling complex in the first 72 h following SAH in the temporal cortex of mice in vivo. A – D Representative Western blot band and densitometric quantification of the time-dependent expression of IP3R1, GRP75, and VDAC1 in the temporal cortex of SAH mice models in the initial 72 h. The expression of the IP3R1–GRP75–VDAC1 complex was upregulated. ns P > 0.05, * P < 0.05. Values were presented as the mean ± SD from four independent experiments ( N = 4). Samples derived from the same experiment, and the gels were processed in parallel. Protein expression levels were normalized to GAPDH as an internal control. Individual gels were run for the separate proteins when the loading protein amount was consistent. E – F Conventional immunofluorescence of PTP1B, Calnexin, VDAC1, and DAPI in the bilateral temporal cortex and CA3. The subsequent MAM junction variation was detected by the conventional immunofluorescence colocalization of calnexin and VDAC1 in vivo. Scale bar = 50 μm, N = 4 per group. G – I The outline and the proportion of surviving neurons induced by SAH in the bilateral temporal cortex and CA3 region. Scale bar = 50 μm, N = 4 per group. J The TEM showed dilated rough ER fragments and swollen mitochondria in the bilateral temporal cortex in each group. Images were acquired at a magnification of 20,000 × , Scale bar = 250 nm. An unpaired Student’s t test was used for comparisons between two groups, whereas ANOVA followed by Tukey’s post hoc test was used for comparisons among multiple groups

Article Snippet: The PVDF membrane was blocked in 5% skim milk for 90 min at room temperature, followed by incubation with the primary antibodies against PTP1B (1:2000 Proteintech, catalog number 11334–1-AP), IP3R1 (1:2000, Abcam, catalog number ab264281), CHOP (1:1000, Proteintech, catalog number 15204–1-AP), Bcl-2 (1:2000, Affinity, lot# 70g9181), Bax (1:2000, Proteintech, catalog number 50599–2-2 g), VDAC1 (1:2000, Proteintech, catalog number 10866–1-AP), AKT (1:1000 Proteintech, catalog number 10176–2-AP), p-AKT (1:1000 Proteintech, catalog number 80455–1-RR), GRP75 (1:1000, Bio-Techne, catalog number MAB3584, ATF4 (1:2000, Proteintech, catalog number 10835–1-AP, p- eIF2a (1:500, Affinity, catalog number AF3087), eIF2a (1:1000, Proteintech, catalog number 11170–1-AP), and CC3 (1:1000, Bioss, Boston, MA, USA lot: BJ03319208), respectively.

Techniques: In Vivo, Western Blot, Expressing, Derivative Assay, Control, Immunofluorescence

Journal: Molecular Neurobiology

Article Title: Metformin Ameliorates Early Brain Injury After Subarachnoid Hemorrhage Via Improving Endoplasmic Reticulum Stress and Mitochondrial Stress-Mediated Ca 2+ Imbalance

doi: 10.1007/s12035-025-05558-1

Figure Lengend Snippet: Met significantly suppressed Ca2 + transfer from the ER to the mitochondria through the PTP1B/AKT following SAH 24 h in the temporal cortex of mice in vivo. A – G To determine the regulatory role of Met in ER stress and mitochondrial signaling, we employed lentivirus-mediated modulation in a 24-h SAH model and confirmed the effects by Western blot analysis. Representative Western blot band and densitometric quantification of the time-dependent expression of CHOP, ATF4, PTP1B, p-eIF2a, eIF2a, p-AKT, and AKT following SAH 24 h in vivo. ns P > 0.05, * P < 0.05. Values were presented as the mean ± SD from four independent experiments ( N = 4). Samples derived from the same experiment, and the gels were processed in parallel. Protein expression levels were normalized to GAPDH as an internal control. Individual gels were run for the separate proteins when the loading protein amount was consistent. H The effect of Met on the concentration of Ca 2+ in the temporal cortex of SAH groups. ns P > 0.05, * P < 0.05. Values were presented as the mean ± SD from four independent experiments ( N = 4). I – L Representative Western blot band and densitometric quantification of the time-dependent expression of IP3R1, GRP75, and VDAC1 in vivo. ns P > 0.05, * P < 0.05. Values were presented as the mean ± SD from four independent experiments ( N = 4). M , N The effect of Met on the conventional immunofluorescence of PTP1B, calnexin, VDAC1, and DAPI in the bilateral temporal cortex and CA3. The subsequent MAM junction variation was detected by the conventional immunofluorescence colocalization of calnexin and VDAC1 in vivo. Scale bar = 50 μm, N = 4 per group. An unpaired Student’s t test was used for comparisons between two groups, whereas ANOVA followed by Tukey’s post hoc test was used for comparisons among multiple groups

Article Snippet: The PVDF membrane was blocked in 5% skim milk for 90 min at room temperature, followed by incubation with the primary antibodies against PTP1B (1:2000 Proteintech, catalog number 11334–1-AP), IP3R1 (1:2000, Abcam, catalog number ab264281), CHOP (1:1000, Proteintech, catalog number 15204–1-AP), Bcl-2 (1:2000, Affinity, lot# 70g9181), Bax (1:2000, Proteintech, catalog number 50599–2-2 g), VDAC1 (1:2000, Proteintech, catalog number 10866–1-AP), AKT (1:1000 Proteintech, catalog number 10176–2-AP), p-AKT (1:1000 Proteintech, catalog number 80455–1-RR), GRP75 (1:1000, Bio-Techne, catalog number MAB3584, ATF4 (1:2000, Proteintech, catalog number 10835–1-AP, p- eIF2a (1:500, Affinity, catalog number AF3087), eIF2a (1:1000, Proteintech, catalog number 11170–1-AP), and CC3 (1:1000, Bioss, Boston, MA, USA lot: BJ03319208), respectively.

Techniques: In Vivo, Western Blot, Expressing, Derivative Assay, Control, Concentration Assay, Immunofluorescence

Journal: Molecular Neurobiology

Article Title: Metformin Ameliorates Early Brain Injury After Subarachnoid Hemorrhage Via Improving Endoplasmic Reticulum Stress and Mitochondrial Stress-Mediated Ca 2+ Imbalance

doi: 10.1007/s12035-025-05558-1

Figure Lengend Snippet: Met significantly suppressed Ca 2+ transfer from the ER to the mitochondria through the PTP1B/AKT following SAH 24 h in vitro. A – K Our vitro data demonstrate that PTP1B plays a critical role in integrating ER stress with mitochondrial dysfunction following SAH 24 h. Representative Western blot band and densitometric quantification of PTP1B, p-AKT, AKT, p-eIF2a, eIF2a, IP3R1, GRP75, VDAC1, ATF4, and CHOP in primary neurons induced by the OxyHb. The expression of the IP3R1–GRP75–VDAC1 complex was upregulated accordingly. ns P > 0.05, * P < 0.05. Values were presented as the mean ± SD from four independent experiments ( N = 4). Samples derived from the same experiment, and the gels were processed in parallel. Protein expression levels were normalized to GAPDH as an internal control. Individual gels were run for the separate proteins when the loading protein amount was consistent. L Conventional immunofluorescence confirmed the colocalization of NEUN, MAP-2, and DAPI in primary neurons. Scale bar = 50 μm. M – R The effect of Met on the apoptosis of SAH in vitro. Representative Western blot bands of Bcl-2, Bax, CC3, and flow cytometry were used to evaluate the effect of Met on apoptosis in vivo. ns P > 0.05, * P < 0.05. Values were presented as the mean ± SD from four independent experiments ( N = 4). Protein expression levels were normalized to GAPDH as an internal control. S The conventional immunofluorescence staining showed that the colocalization of intracellular Ca 2+ concentration increased significantly in the OxyHb24H group in primary neurons. Scale bar = 50 μm. T The conventional immunofluorescence confirmed the colocalization of PTP1B, calnexin, VDAC1, and DAPI in primary neurons after the intervention of lentivirus and inhibitors. To assess the effect of Met on MAM integrity, we performed immunofluorescence co-localization analysis of the ER marker calnexin and the mitochondrial marker VDAC1 in treated primary neurons. Scale bar = 50 μm. An unpaired Student’s t test was used for comparisons between two groups, whereas ANOVA followed by Tukey’s post hoc test was used for comparisons among multiple groups

Article Snippet: The PVDF membrane was blocked in 5% skim milk for 90 min at room temperature, followed by incubation with the primary antibodies against PTP1B (1:2000 Proteintech, catalog number 11334–1-AP), IP3R1 (1:2000, Abcam, catalog number ab264281), CHOP (1:1000, Proteintech, catalog number 15204–1-AP), Bcl-2 (1:2000, Affinity, lot# 70g9181), Bax (1:2000, Proteintech, catalog number 50599–2-2 g), VDAC1 (1:2000, Proteintech, catalog number 10866–1-AP), AKT (1:1000 Proteintech, catalog number 10176–2-AP), p-AKT (1:1000 Proteintech, catalog number 80455–1-RR), GRP75 (1:1000, Bio-Techne, catalog number MAB3584, ATF4 (1:2000, Proteintech, catalog number 10835–1-AP, p- eIF2a (1:500, Affinity, catalog number AF3087), eIF2a (1:1000, Proteintech, catalog number 11170–1-AP), and CC3 (1:1000, Bioss, Boston, MA, USA lot: BJ03319208), respectively.

Techniques: In Vitro, Western Blot, Expressing, Derivative Assay, Control, Immunofluorescence, Flow Cytometry, In Vivo, Staining, Concentration Assay, Marker

Journal: Molecular Neurobiology

Article Title: Metformin Ameliorates Early Brain Injury After Subarachnoid Hemorrhage Via Improving Endoplasmic Reticulum Stress and Mitochondrial Stress-Mediated Ca 2+ Imbalance

doi: 10.1007/s12035-025-05558-1

Figure Lengend Snippet: Metformin ameliorates early brain injury after subarachnoid hemorrhage via improving ERS and MS-mediated Ca 2+ imbalance. The IP3R1–GRP75–VDAC1 complex regulates MAM and plays an imperative role in EBI. The treatment of Met ameliorates ER stress, MS, and Ca 2+ overload, further improving neuroapoptosis and alleviating the EBI of SAH

Article Snippet: The PVDF membrane was blocked in 5% skim milk for 90 min at room temperature, followed by incubation with the primary antibodies against PTP1B (1:2000 Proteintech, catalog number 11334–1-AP), IP3R1 (1:2000, Abcam, catalog number ab264281), CHOP (1:1000, Proteintech, catalog number 15204–1-AP), Bcl-2 (1:2000, Affinity, lot# 70g9181), Bax (1:2000, Proteintech, catalog number 50599–2-2 g), VDAC1 (1:2000, Proteintech, catalog number 10866–1-AP), AKT (1:1000 Proteintech, catalog number 10176–2-AP), p-AKT (1:1000 Proteintech, catalog number 80455–1-RR), GRP75 (1:1000, Bio-Techne, catalog number MAB3584, ATF4 (1:2000, Proteintech, catalog number 10835–1-AP, p- eIF2a (1:500, Affinity, catalog number AF3087), eIF2a (1:1000, Proteintech, catalog number 11170–1-AP), and CC3 (1:1000, Bioss, Boston, MA, USA lot: BJ03319208), respectively.

Techniques: