ero1  (Proteintech)

Journal: bioRxiv

Article Title: Chronic ER Stress Disrupts Mitochondrial-Associated ER Membrane Integrity in Corneal Endothelial Cells

doi: 10.64898/2026.01.09.698664

Figure Lengend Snippet: (A) Description of the method of subcellular fractionation for mitochondrial-associated ER membrane (MAMs), endoplasmic reticulum (ER), and crude mitochondria (CM) in HCEnC-21T cell line. (B) Representative Western blot data confirming subcellular fraction markers PDZD8 (MAM marker), Ero1-Lα (ER marker), VDAC, and cytochrome C (mitochondrial markers), GAPDH (loading control) in normal 21T and diseased F35T cell line. (C) Immunostained images of MAM tracker-Green transfected 21T and F35T cells showing no significant difference in intensity, indicating no disruption of ER-mitochondrial interactions/ MAMs in both cell lines at baseline (non-stressed).

Article Snippet: The following antibodies were used: PDZD8 (cat no. PA5-46771, Invitrogen), Ero1-Lα (cat no. 3264, Cell Signaling Technology), VDAC (cat no. 4661, Cell Signaling Technology), cytochrome C (cat no. 4272, Cell Signaling Technology), PERK (cat no. 3192, Cell Signaling Technology), Parkin (cat no. 4211, Cell Signaling Technology), GAPDH (cat no. 97166, Cell Signaling Technology), actin (cat no. 3700, Cell Signaling Technology), HRP-linked anti-mouse (cat no. 7076, Cell Signaling Technology) and anti-rabbit (cat no. 7074, Cell Signaling Technology) secondary antibodies.

Techniques: Fractionation, Membrane, Western Blot, Marker, Control, Transfection, Disruption

Journal: bioRxiv

Article Title: Chronic ER Stress Disrupts Mitochondrial-Associated ER Membrane Integrity in Corneal Endothelial Cells

doi: 10.64898/2026.01.09.698664

Figure Lengend Snippet: (A) Representative Western blots of subcellular fractions of 21T and F35T cell line at baseline (untreated), probing for PERK, Parkin and GAPDH (B) Bar graph demonstrating increased Parkin expression in the MAM fraction of diseased F35T corneal endothelial cells compared to healthy 21T cells at baseline (n = 3, *p < 0.05, unpaired t-test). (C) Representative Western blots confirming subcellular fraction markers PDZD8 (MAM marker), Ero1-Lα (ER marker), VDAC, and cytochrome C (mitochondrial markers), GAPDH (loading control) in normal 21T cells after treatment with control DMSO (0.02%) or ER stressor tunicamycin (1 µg/mL) at 24 hrs. (D) Representative Western blots showing increased expression of PERK and Parkin proteins in the MAM fraction in 21T cells after ER stressor tunicamycin (1 µg/mL) at 24 hours compared to control DMSO (0.02%) (B) Bar graph also demonstrating an increase in expression level of PERK and Parkin under the TUN condition compared to control (n = 4, *p < 0.05, unpaired t-test).

Article Snippet: The following antibodies were used: PDZD8 (cat no. PA5-46771, Invitrogen), Ero1-Lα (cat no. 3264, Cell Signaling Technology), VDAC (cat no. 4661, Cell Signaling Technology), cytochrome C (cat no. 4272, Cell Signaling Technology), PERK (cat no. 3192, Cell Signaling Technology), Parkin (cat no. 4211, Cell Signaling Technology), GAPDH (cat no. 97166, Cell Signaling Technology), actin (cat no. 3700, Cell Signaling Technology), HRP-linked anti-mouse (cat no. 7076, Cell Signaling Technology) and anti-rabbit (cat no. 7074, Cell Signaling Technology) secondary antibodies.

Techniques: Western Blot, Expressing, Marker, Control