ma  (MedChemExpress)

Journal: iScience

Article Title: MSMO1 promotes chemotherapy resistance through modulation of T-MAS metabolism via PERK/elF2α/ATF4/CHOP pathway

doi: 10.1016/j.isci.2026.114790

Figure Lengend Snippet: T-MAS induces ER stress and activates PERK/eIF2α/ATF4/CHOP pathway (A) Diagram of the cholesterol metabolism pathway, using siRNA to inhibit TM7SF2 to reduce the cellular levels of T-MAS. (B and C) Western blotting analysis showing the expression of BIP, PERK, p -eIF2α, ATF4, CHOP in MSMO1-KD and control cells with or without siRNA interfering with TM7SF2. (D) Diagram of the cholesterol metabolism pathway, using siRNA to inhibit CYP51A1 to reduce the cellular levels of FF-MAS and downstream metabolites. (E) Western blotting analysis showing the expression of BIP, PERK, p -eIF2α, ATF4, CHOP in wild-type SK-BR-3 cells with or without the pretreatment of 100 nM Tg for 3 h and siRNA interfering with CYP51A1. (F) Western blotting images showing the expression of BIP, PERK, p -eIF2α, ATF4, and CHOP in MSMO1-KD and control MDA-MB-231 cells with or without siRNA interfering with CYP51A1. (G and I) Schematic view of silencing TM7SF2 through siRNA to inhibit the transformation of FF-MAS to T-MAS, followed by the administration of FF-MAS or T-MAS to the cells. (H and J) Western blotting analysis showing the expression of BIP, PERK, p -eIF2α, ATF4, and CHOP with or without siRNA interfering with TM7SF2 and 1 μg/mL FF-MAS(or 1 μg/mL T-MAS) treatment of 48 h.

Article Snippet: FF-MAS , Avanti , Cat#: 700077P.

Techniques: Western Blot, Expressing, Control, Transformation Assay

Journal: iScience

Article Title: MSMO1 promotes chemotherapy resistance through modulation of T-MAS metabolism via PERK/elF2α/ATF4/CHOP pathway

doi: 10.1016/j.isci.2026.114790

Figure Lengend Snippet: MSMO1 modulates the relative content of T-MAS to regulate endoplasmic reticulum stress (A) Schematic diagram of cholesterol metabolic pathways, starting with squalene. (B) Heatmap showing the cholesterol metabolites with the most statistically significant differences between MSMO1-KD MDA-MB-231 and control cells. (C) Volcano plots showing differential metabolites between MSMO1-KD and control cells. T-MAS exhibited a significant increase (Log 2 (fold change) = 5.66 p < 0.001) in MSMO1-KD cells. (D) Representative images of thioflavin T (ThT) staining assay with or without the treatment of 1 mM 4-PBA for 3 h and 1 μg/mL T-MAS for 48 h. Scale bars, 10 μm. (E) Western blotting analysis showing the expression of BIP, PERK, p -eIF2α, ATF4, and CHOP in MDA-MB-231 cells with or without the pretreatment of 1 μM PERKi for 3 h and the treatment of 1 μg/mL T-MAS for 48 h. (F and G) IC50 curves of MSMO1 overexpressing and MSMO1-KD cells and the corresponding control cells for Tg with or without the treatment of 1 μg/mL T-MAS. Data are represented as mean ± SEM.

Article Snippet: T-MAS , Avanti , Cat#: 700073P.

Techniques: Control, Staining, Western Blot, Expressing

Journal: iScience

Article Title: MSMO1 promotes chemotherapy resistance through modulation of T-MAS metabolism via PERK/elF2α/ATF4/CHOP pathway

doi: 10.1016/j.isci.2026.114790

Figure Lengend Snippet: T-MAS induces ER stress and activates PERK/eIF2α/ATF4/CHOP pathway (A) Diagram of the cholesterol metabolism pathway, using siRNA to inhibit TM7SF2 to reduce the cellular levels of T-MAS. (B and C) Western blotting analysis showing the expression of BIP, PERK, p -eIF2α, ATF4, CHOP in MSMO1-KD and control cells with or without siRNA interfering with TM7SF2. (D) Diagram of the cholesterol metabolism pathway, using siRNA to inhibit CYP51A1 to reduce the cellular levels of FF-MAS and downstream metabolites. (E) Western blotting analysis showing the expression of BIP, PERK, p -eIF2α, ATF4, CHOP in wild-type SK-BR-3 cells with or without the pretreatment of 100 nM Tg for 3 h and siRNA interfering with CYP51A1. (F) Western blotting images showing the expression of BIP, PERK, p -eIF2α, ATF4, and CHOP in MSMO1-KD and control MDA-MB-231 cells with or without siRNA interfering with CYP51A1. (G and I) Schematic view of silencing TM7SF2 through siRNA to inhibit the transformation of FF-MAS to T-MAS, followed by the administration of FF-MAS or T-MAS to the cells. (H and J) Western blotting analysis showing the expression of BIP, PERK, p -eIF2α, ATF4, and CHOP with or without siRNA interfering with TM7SF2 and 1 μg/mL FF-MAS(or 1 μg/mL T-MAS) treatment of 48 h.

Article Snippet: T-MAS , Avanti , Cat#: 700073P.

Techniques: Western Blot, Expressing, Control, Transformation Assay

Journal: iScience

Article Title: MSMO1 promotes chemotherapy resistance through modulation of T-MAS metabolism via PERK/elF2α/ATF4/CHOP pathway

doi: 10.1016/j.isci.2026.114790

Figure Lengend Snippet: T-MAS enhances the chemosensitivity of breast cancer organoids (A) Representative electron microscope (EM) images of untreated, 1 μg/mL T-MAS treated, 500 nM PTX treated, and 500 nM PTX plus 1 μg/mL T-MAS treated cells. Arrowheads indicate the ER. 120 ER widths were measured in each group. x5k scale bars, 5 μm. x25k scale bars, 1 μm. (B) Schematic view of sample collection ( n = 7). Tissue specimens from the primary tumor were collected by core needle biopsy and then induced into organoids for drug sensitivity testing. Plasma exosomes were further extracted from the blood sample. (C) Correlation analysis between primary tumor site and plasma exosome MSMO1 expression level (R = 0.86, p = 0.013). (D) Representative images of fluorescence microscopy show Live (AO = green)/dead (PI = red) analysis of breast cancer organoids following treatment of 500 nM PTX or 200 μM CBP or in combination with 1 μg/mL T-MAS. Scale bars, 20 μm. (E) Quantification results of organoid live/dead analysis. Data are represented as mean ± SEM. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗∗ p < 0.0001.

Article Snippet: T-MAS , Avanti , Cat#: 700073P.

Techniques: Microscopy, Clinical Proteomics, Expressing, Fluorescence

Journal: Journal of Orthopaedic Translation

Article Title: HIF1A transcriptionally activates CDKN1A to drive ferroptosis in skeletal muscle ischaemia-reperfusion injury

doi: 10.1016/j.jot.2026.101055

Figure Lengend Snippet: Ferroptosis drives skeletal muscle I/R injury and is rescued by Ferrostatin-1 in vivo and in vitro . (A) Representative HE-stained photomicrographs of gastrocnemius muscle sections from Sham, I/R, Sham + Fer-1, and I/R + Fer-1 experimental groups. Neutrophil infiltration was observed following reperfusion, and this infiltration was alleviated with Fer-1 treatment. (B) Histological injury score. (C) Skeletal muscle wet/dry weight ratio. (D) GSH level in skeletal muscle tissues. (E) MDA level in skeletal muscle tissues. (F) ROS level in skeletal muscle tissues. (G) Iron level in skeletal muscle tissues. (H) Infarct ratio. (I) Representative images of TTC-stained skeletal muscle sections of different groups. (J) Representative TEM images of different groups. (K-N) The protein levels of GPX4, ACSL4, PTGS2 in vivo . (O) CCK-8 assay to detect the inhibition rate of C2C12 cells under different H/R durations. (P) C2C12 cells were treated with the ferroptosis inducer Erastin, the ferroptosis inhibitor Fer-1, the apoptosis inhibitor Z-VAD, the autophagy inhibitor 3-MA, and the necroptosis inhibitor Nec-1, and cell inhibition rate was assayed using CCK-8. (Q) GSH level. (R) MDA level. (S) ROS level. (T) Iron level. (U) Representative TEM images of different groups. (V-Y) The protein levels of GPX4, ACSL4, PTGS2 in vitro . Data are expressed as mean ± SD. For in vivo experiments, each group comprised 8-10 animals, with n = 3 randomly selected animals per assay. For in vitro experiments, n = 3 independent experiments. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001; ns, non-significant.

Article Snippet: Concurrently, apoptosis, autophagy, and necroptosis pathways were selectively inhibited through treatment with pan-caspase inhibitor Z-VAD (10 μM, T6013, Topscience, China), autophagy inhibitor 3-MA (5 mM, HY-19312, MCE, China), and necroptosis inhibitor Nec-1 (10 μM, T1847, Topscience, China), respectively.

Techniques: In Vivo, In Vitro, Staining, CCK-8 Assay, Inhibition

Journal: The Journal of Biological Chemistry

Article Title: A GluN2B disease-associated variant promotes the degradation of NMDA receptors via autophagy

doi: 10.1016/j.jbc.2026.111147

Figure Lengend Snippet: The GluN2B subunit undergoes degradation via autophagy . A , inhibition of the proteasome with MG132 (10 μM) and the lysosome with Bafilomycin-A1 (1 μM) for 6 h effect on the GluN2B subunit in HEK293T cells stably expressing recombinant WT or R519Q NMDARs. β-actin serves as the soluble total protein loading control (n = 5). B , effects of R519Q on total GluN2B protein expression levels 48 h post transient transfection with GluN1 and GluN2B constructs at a 1:1 ratio in ATG7 KO HEK293T to express WT or GluN2B_R519Q variant NMDARs. β-actin served as the soluble total protein loading control. (n = 6). C , surface biotinylation assay to monitor the influence of the R519Q DAV on the surface expression of NMDARs expressed in ATG7 KO HEK293T 48 h post transient transfection. Na + /K + ATPase served as a membrane protein loading control (n = 6). D , schematic of macroautophagy pathway, including induction, nucleation and formation of the isolation membrane, elongation, autophagosome formation, and fusion with the lysosome forming the autophagolysosome. Rapamycin is shown to inhibit mTOR, which results in autophagy activation, while 3-MA and Baf-A1 inhibit autophagy at different stages as shown. E , HEK293T cells stably expressing R519Q NMDARs were treated with autophagy activators (SMER28 10 μM, Rapamycin 100 nM) and inhibitors of autophagy (3-MA 50 mM and Baf-A1 20 nM) for 24 h. Changes to total GluN2B expression were monitored via immunoblot and p62 was used as a marker for autophagic flux. β-actin serves as the soluble total protein loading control (n = 3). F , siRNA knockdown of ATG7 effect on R519Q variant GluN2B expression and immunoblot was performed 48 h after knockdown. β-actin served as the soluble total protein loading control (n = 3). Non-targeting (NT) siRNA was used as a control for each condition, and two independent siRNA constructs were used to knockdown gene expression. G , siRNA knockdown of LC3b effects on R519Q variant GluN2B expression. Immunoblot performed 48 h after knockdown. β-actin served as the soluble total protein loading control (n = 3). Non-targeting siRNA was used as a control for each condition, and two independent siRNA constructs were used to knockdown gene expression. LC3-Pool contains two unique siRNAs for each of the LC3a, LC3b, and LC3c isoforms. All data are normalized to the appropriate loading control, and data are presented as mean ± SD. Statistical significance was determined using an unpaired two-tailed Student’s t test between two groups or an analysis of variance (ANOVA) followed by a post hoc Dunnett’s test for comparison in multiple groups. Significance level defined as ns, not significant, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: The 3-methyladenine (3-MA) (#HY-19312), SMER28 (#HY-100200), rapamycin (#HY-10219), and DAPI dihydrochloride (HY-D0814) were obtained from MedChemExpress.

Techniques: Inhibition, Stable Transfection, Expressing, Recombinant, Control, Transfection, Construct, Variant Assay, Surface Biotinylation Assay, Membrane, Isolation, Activation Assay, Western Blot, Marker, Knockdown, Gene Expression, Two Tailed Test, Comparison