anti p drp1 ser616  (Cell Signaling Technology Inc)

Journal: Cell Death & Disease

Article Title: Dual-faced metabolic reprogramming powers proliferation: FGF21 drives lung squamous carcinoma progression via CaMKII-DRP1-mediated mitochondrial fission

doi: 10.1038/s41419-026-09084-3

Figure Lengend Snippet: A Representative Western blotting images showing the expression of mitochondrial fusion and fission proteins in response to FGF21 treatment. The protein levels were normalized to the GAPDH levels. B Representative Western blotting images showing the expression of p-DRP1 Ser616 and DRP1. C Immunofluorescence staining showing the expression level of p-DRP1 Ser616; scale bars, 100 μm. Fiji software was used for image analysis, and the average fluorescence intensity was calculated. D Fractions of LUSC cell lysates, showing the levels of DRP1 in the mitochondrial (Mito) and cytoplasmic (Cyto) fractions. The mitochondrial protein COX-IV and the cytoplasmic protein β-actin served as controls. E Confocal microscopy images showing the levels of mitochondria-bound DRP1 in LUSC cells treated with FGF21. DRP1 and mitochondria were labeled with a mouse anti-DRP1 antibody and Mito Tracker Green, respectively; scale bars, 10 μm (annotation added; original software-generated scale bar displayed a “too long” error due to a display glitch). Pearson’s coefficient indicates colocalization of DRP1 and Mito Tracker Green. The data are shown as the means ± SEMs or SDs; * p < 0.05, ** p < 0.01, *** p < 0.01.

Article Snippet: Anti-p-DRP1 (ser616) , 4494 , Rabbit , CST , WB (1:1000); IF (1:1000).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Software, Analysis, Fluorescence, Confocal, Microscopy, Labeling, Generated

Journal: Cell Death & Disease

Article Title: Dual-faced metabolic reprogramming powers proliferation: FGF21 drives lung squamous carcinoma progression via CaMKII-DRP1-mediated mitochondrial fission

doi: 10.1038/s41419-026-09084-3

Figure Lengend Snippet: A Representative western blotting images showing the protein expression of p-CaMKII (Thr286) in H1703 and H520 cells incubated with FGF21 for different durations. B Representative Western blotting images showing the protein expression of p-CaMKII (Thr286), CaMKII, p-DRP1 (Ser616) and DRP1 in H1703 and H520 cells incubated with FGF21 pretreated with KN93. C Immunofluorescence staining showing the expression and distribution of p-DRP1 (Ser616) and DRP1 in H1703 and H520 cells incubated with FGF21 pretreated with KN93. D Immunofluorescence staining showing the mitochondrial morphology in H1703 and H520 cells incubated with FGF21 pretreated with KN93. E CCK-8 assay showing the viability of H1703 and H520 cells incubated with FGF21 pretreated with or without KN93. F Representative western blotting images showing the expression of key glycolytic pathway components (GLUT1, LDHA) in DRP1-silenced LUSC cells with or without FGF21 stimulation. GAPDH served as a loading control. The data are shown as the means ± SEMs or SDs; * p < 0.05, ** p < 0.01, *** p < 0.01, **** p < 0.01.

Article Snippet: Anti-p-DRP1 (ser616) , 4494 , Rabbit , CST , WB (1:1000); IF (1:1000).

Techniques: Western Blot, Expressing, Incubation, Immunofluorescence, Staining, Morphology, CCK-8 Assay, Control

Journal: Cell Death & Disease

Article Title: Dual-faced metabolic reprogramming powers proliferation: FGF21 drives lung squamous carcinoma progression via CaMKII-DRP1-mediated mitochondrial fission

doi: 10.1038/s41419-026-09084-3

Figure Lengend Snippet: List of primary antibodies, secondary antibodies, and nuclear dyes used in this study.

Article Snippet: Anti-p-DRP1 (ser616) , 4494 , Rabbit , CST , WB (1:1000); IF (1:1000).

Techniques:

Journal: bioRxiv

Article Title: Retrosplenial cortex vulnerability links severe hypoglycemia to cognitive impairment through neuron–microglia crosstalk

doi: 10.64898/2026.03.27.714654

Figure Lengend Snippet: (a) Experimental design for mdivi-1 and minocycline treatment. (b) Blood glucose levels. (c) Representative images of NeuN and 4-HNE co-staining in the RSC. Scale bars, 50 μm. White arrows indicate 4-HNE + cells co-localized with NeuN + cells. (d) Quantification of the percentage of 4-HNE + cells co-localized with NeuN + cells ( n = 5 mice per group). (e) Representative images of Iba-1 staining in the RSC. Scale bars, 100 μm. (f) Quantification of Iba-1 + cells normalized by the area size of 0.64 mm 2 ( n = 5 mice per group). (g-i) Representative Z-stack images of Iba-1 + microglia in the RSC (g). Scale bars, 10 μm. The branched paths of microglia were selected based on the captured images. Quantification of microglial branch number (h) and path length (i) ( n = 27-32 independent microglia). Each dot represents the quantification of a single microglial cell from n = 5 biologically independent samples in each group. (j) Representative images in the RSC sections stained with NeuN and pDrp1(S616). White arrows indicate pDrp1 + cells co-localized with NeuN + cells. Scale bars, 50 μm. (k) Quantification of the percentage of pDrp1 + cells co-localized with NeuN + cells ( n = 5 mice per group). Data are presented as means ± SEM and were analyzed by one-way ANOVA followed by Dunnett’s multiple comparisons test. #P < 0.05; ##P < 0.01; ###P < 0.001.

Article Snippet: After being washed in PBS, the brain sections were blocked with 0.3% Triton X-100 and 5% donkey serum in PBS for 1 hour at room temperature, followed by 24 hours of incubation at 4°C with primary antibodies [anti-4-HNE antiserum (Alpha Diagnostic, HNE11S, 1:200), anti-MAP2 (Millipore Sigma, AB5622, 1:200), anti-Iba-1 (Wako Chemicals, #01919741, 1:500; Abcam, ab5076, 1:500), anti-NeuN (EMD Millipore, MAB377, 1:200), anti-GFAP (Invitrogen, MA5-12023, 1:400), and anti-pDrp1 (S616) (Cell Signaling Technology, #3455, 1:100)].

Techniques: Staining

Journal: bioRxiv

Article Title: Retrosplenial cortex vulnerability links severe hypoglycemia to cognitive impairment through neuron–microglia crosstalk

doi: 10.64898/2026.03.27.714654

Figure Lengend Snippet: (a) Experimental design for IL-1ra ICV injection. (b) Blood glucose levels. (c) Representative images of the RSC tissues with NeuN and 4-HNE co-staining. White arrows indicate 4-HNE + cells co-localized with NeuN + cells. Scale bars, 20 μm. (d) Quantification of the percentage of 4-HNE + cells co-localized with NeuN + cells ( n = 4 mice per group). (e) Representative images of the RSC tissues with pDrp1(S616) and NeuN co-staining. White arrows indicate pDrp1 + cells co-localized with NeuN + cells. Scale bars, 20 μm. (f) Quantification of the percentage of pDrp1 + cells co-localized with NeuN + cells ( n = 4 mice per group). (g) Representative images of Iba-1 staining. Scale bars, 100 μm. (h) Quantification of Iba-1 + cells normalized by the area of 0.64 mm 2 ( n = 4 mice per group). (i) Representative Z-stack images in the RSC sections with Iba-1 staining. Scale bars, 20 μm. (j and k) Quantification of the number of branches (j) and path length (k) ( n = 19-26 independent microglia). Each dot represents the quantification of a single microglial cell from n = 4 biologically independent samples in each group. (l) Experimental design for IL-1ra pretreatment in BV-2 or SH-SY5Y cells. (m) Representative images of SH-SY5Y cells with cleaved caspase-3 staining. Scale bars, 50 μm. (n) Quantification of cleaved caspase-3 + cells as a percentage of DAPI + cells ( n = 5 or 6 independent samples per group). (o) Representative Z-stack images of SH-SY5Y cells with MitoTracker staining. Scale bars, 10 μm. (p) Quantification of mitochondrial length ( n = 5 independent samples per group). Data are presented as means ± SEM and were analyzed by one-way ANOVA followed by Dunnett’s multiple comparisons test. #P < 0.05; ###P < 0.001.

Article Snippet: After being washed in PBS, the brain sections were blocked with 0.3% Triton X-100 and 5% donkey serum in PBS for 1 hour at room temperature, followed by 24 hours of incubation at 4°C with primary antibodies [anti-4-HNE antiserum (Alpha Diagnostic, HNE11S, 1:200), anti-MAP2 (Millipore Sigma, AB5622, 1:200), anti-Iba-1 (Wako Chemicals, #01919741, 1:500; Abcam, ab5076, 1:500), anti-NeuN (EMD Millipore, MAB377, 1:200), anti-GFAP (Invitrogen, MA5-12023, 1:400), and anti-pDrp1 (S616) (Cell Signaling Technology, #3455, 1:100)].

Techniques: Injection, Staining

Journal: bioRxiv

Article Title: Retrosplenial cortex vulnerability links severe hypoglycemia to cognitive impairment through neuron–microglia crosstalk

doi: 10.64898/2026.03.27.714654

Figure Lengend Snippet: (a) Schematic of Drp1-dependent mitochondrial fission. (b) Schematic of mitochondria isolation. (c and d) Immunoblot analysis from isolated mitochondria fraction of the RSC tissues ( n = 4 or 5 mice). Data are presented as means ± SEM and were analyzed by one-way ANOVA followed by Tukey’s multiple comparisons test. **P < 0.01; ***P < 0.001. (e) Experimental design of the study. Dnm1l ( n = 9 mice) or Il1b ( n = 10 mice) siRNA was injected in the right RSC while control siRNA was injected in the contralateral RSC. (f) Blood glucose levels. (g-n) Immunoblot analysis from the RSC tissues with antibody against mitochondrial fission proteins (pDrp1(S616) and Drp1) and IL-1β on day 1 after hypoglycemia induction ( n = 4 or 5 mice per group). (o-r) Immunohistochemistry analysis from the RSC tissues with NeuN and 4-HNE co-staining on day 7 after hypoglycemia induction ( n = 5 mice per group). White arrows indicate 4-HNE + cells co-localized with NeuN + cells. Scale bars, 20 μm. Each dot represents a single RSC site from n = 4 or 5 mice injected unilaterally with Dnm1l or Il1b siRNA and contralaterally with control siRNA. Paired t-tests were used for statistical comparisons between hemispheres. Data are presented as means ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: After being washed in PBS, the brain sections were blocked with 0.3% Triton X-100 and 5% donkey serum in PBS for 1 hour at room temperature, followed by 24 hours of incubation at 4°C with primary antibodies [anti-4-HNE antiserum (Alpha Diagnostic, HNE11S, 1:200), anti-MAP2 (Millipore Sigma, AB5622, 1:200), anti-Iba-1 (Wako Chemicals, #01919741, 1:500; Abcam, ab5076, 1:500), anti-NeuN (EMD Millipore, MAB377, 1:200), anti-GFAP (Invitrogen, MA5-12023, 1:400), and anti-pDrp1 (S616) (Cell Signaling Technology, #3455, 1:100)].

Techniques: Isolation, Western Blot, Injection, Control, Immunohistochemistry, Staining