fdft1  (Proteintech)

Journal: iScience

Article Title: APOE4 drives widespread changes to the hepatic proteome and alters metabolic function

doi: 10.1016/j.isci.2026.115035

Figure Lengend Snippet: Comparative proteomic analysis of whole mouse livers and iHLCs (A) Venn diagram depicting overlap of significant DE proteins in APOE4 vs. APOE3 across whole mouse livers and iHLCs (batch 1 and 2). (B) Gene Ontology Biological Process (GO-BP) and KEGG pathway analysis of the 70 proteins shared across mouse liver and iHLCs, showing protein counts per category. (C) Heatmap of proteins consistently upregulated or downregulated in APOE4 vs. APOE3 across whole mouse liver and iHLCs (batch 1 and 2). (D–E) Western blot analysis of CYP27A1, FDFT1, and PGAM1 in APOE4 and APOE3 whole mouse livers, with relative densitometry quantification. n = 16 per group. (F–G) Western blot analysis of CYP27A1, FDFT1, and PGAM1 in APOE4 and APOE3 isogenic iHLCs, with relative densitometry quantification. n = 6 per group. (E and G) Statistical significance was determined by unpaired t test. Data are shown as mean ± SD. ∗ p < 0.05.

Article Snippet: Primary antibodies included ALB (Abcam, ab207327), APOE (Abcam, ab183597), ASGR1 (Abcam, ab254261), CS (Cell Signaling, #14309), CYP27A1 (Abcam, ab126785), FDFT1 (Proteintech, 83020-1-RR), HDAC1 (Cell Signaling, #34589), HK1 (Cell Signaling, #2024), HNF4A (Abcam, ab92378), NANOG (Abcam, ab109250), OCT4 (Abcam, ab181557), PGAM1 (Cell Signaling, #12098), and SOX17 (Abcam, ab224637).

Techniques: Western Blot

Journal: Advanced Science

Article Title: Schwann Cell Synthesized Cholesterol Orchestrates Peripheral Nerve Regeneration via Structural and IGF1‐Dependent Signaling Mechanisms

doi: 10.1002/advs.202520323

Figure Lengend Snippet: The expression pattern of cholesterol synthesis‐related genes in SCs following PNI. (A) Subpopulation analysis based on reprocessed single‐cell RNA‐sequencing data ( GSE216665 ). (B,C) Altered expression and cellular localization patterns of key enzymes in the cholesterol synthesis pathways at 3, 12 and 60 dpi. (D) The expression dynamics of FDFT1 in SCs at 3, 12 and 60 dpi. (E) Western blot analysis of FDFT1 protein levels in the sciatic nerve at 3, 7, 14 and 28 dpi ( n = 6–8). (F) Double immunofluorescence staining and quantification (e1) for FDFT1 (red) and SCs marker S100 (green) in transverse sections of the injured nerves at 3, 7, 14 and 28 dpi. Scale bar = 20 µm, zoom in, 5 µm, ( n = 5). Data are presented as mean ± SD. Statistical analysis was performed using one‐way ANOVA for multiple comparisons, “ns” indicating no significance, * p < 0.05.

Article Snippet: The Fdft1 flox/flox mice, Plp CreERT2 mice and Dhh Cre mice were obtained from Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Expressing, Single Cell, RNA Sequencing, Western Blot, Double Immunofluorescence Staining, Marker

Journal: Advanced Science

Article Title: Schwann Cell Synthesized Cholesterol Orchestrates Peripheral Nerve Regeneration via Structural and IGF1‐Dependent Signaling Mechanisms

doi: 10.1002/advs.202520323

Figure Lengend Snippet: Generation and identification of SC‐specific FDFT1 knockout mice. (A,B) Breeding strategies of the two lines of conditional knockout mice. (C,D) Representative genotyping images of icKO ( Fdft1 flox/flox ; Plp CreERT2 ) and cKO ( Fdft1 flox/flox ; Dhh Cre ) mice. (E,F) FDFT1 deletion efficiency in adult icKO mice, as shown by double immunofluorescence staining for FDFT1 (red) and S100 (green), and western blotting analysis in (F) ( n = 6). Scale bar = 20 µm, zoom in, 5 µm. (G,H) Immunofluorescence and Western blot analyses (H) confirm the FDFT1‐knockout efficiency in primary SCs isolated from cKO neonatal mice. Scale bar = 50 µm, zoom in, 10 µm ( n = 4). I) Filipin III staining (blue) and Phalloidin staining (red) visualize the cholesterol levels in the primary cultured SCs. Scale bar = 100 µm. All data are presented as mean ± SD. Two‐tailed Student's t ‐test, “ns” indicating no significance, * p < 0.05.

Article Snippet: The Fdft1 flox/flox mice, Plp CreERT2 mice and Dhh Cre mice were obtained from Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Knock-Out, Double Immunofluorescence Staining, Western Blot, Immunofluorescence, Isolation, Staining, Cell Culture, Two Tailed Test

Journal: Advanced Science

Article Title: Schwann Cell Synthesized Cholesterol Orchestrates Peripheral Nerve Regeneration via Structural and IGF1‐Dependent Signaling Mechanisms

doi: 10.1002/advs.202520323

Figure Lengend Snippet: FDFT1 deficiency in SCs impairs axonal regeneration and remyelination following sciatic nerve crush injury. (A) Immunostaining and quantifications (a1) show the GAP43 + regenerating axons in longitudinal sections of the injured nerves at 3 dpi. Scale bars = 200 µm, ( n = 5). (B) Immunostaining displays GAP43 + axons in cross sections 3 mm distal to the injury site at 3 dpi, quantification shown in (b1). Scale bar = 100 µm, zoom in, 10 µm, ( n = 5). (C) Western blot analysis and quantification (c1) of GAP43 protein levels in the distal nerve trunks at 3 dpi ( n = 8). (D) Immunostaining and fluorescence intensity quantification (d1) of P0 (red) and NF200 (green) on the transverse sections of the nerve at 3 mm distal to the lesion site at 28 dpi. Scale bar = 20 µm, zoom in, 5 µm, ( n = 5). (E) Western blotting and protein levels (e1) of MBP and NF200 in the distal trunk of the injured nerve at 28 dpi ( n = 7–8). (F) Transmission electron microscopy (TEM) analysis of the regenerated nerve 3 mm distal to the lesion at 28 dpi, showing axon area (f1), myelin thickness(f2), the number of myelinated axons (f3) and G‐ratio (f4), ( n = 4). Data are presented as mean ± SD. Two‐tailed Student's t ‐test, “ns” indicating no significance, * p < 0.05.

Article Snippet: The Fdft1 flox/flox mice, Plp CreERT2 mice and Dhh Cre mice were obtained from Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Immunostaining, Western Blot, Fluorescence, Transmission Assay, Electron Microscopy, Two Tailed Test

Journal: Advanced Science

Article Title: Schwann Cell Synthesized Cholesterol Orchestrates Peripheral Nerve Regeneration via Structural and IGF1‐Dependent Signaling Mechanisms

doi: 10.1002/advs.202520323

Figure Lengend Snippet: SC‐specific FDFT1 knockout hinders morphological and functional recovery after sciatic nerve injury at 28 dpi. (A) The hindlimb extension and paw area (a1) during suspension. Scale bar = 5 mm, ( n = 4). (B,C) Representative footprints image, and the SFI scores shown in (C). Scale bar = 1 cm, ( n = 8). (D) Motor coordination assessed by rota‐rod test at accelerating speed from 5 rpm to 40 rpm over 120 s. Quantification of time to fall (d1), ( n = 4–8). (E) Respective CMAP images and the quantification of latency (e1) and amplitude (e2), ( n = 8). (F) Gross morphology and wet weight ratio (f1) of the gastrocnemius muscle, ( n = 8). (G) HE staining and fiber cross‐sectional area analyses (g1) of the gastrocnemius muscle. Scale bar = 100 µm, ( n = 4–5). Data are presented as mean ± SD. Two‐way ANOVA, “ns” indicating no significance, * p < 0.05.

Article Snippet: The Fdft1 flox/flox mice, Plp CreERT2 mice and Dhh Cre mice were obtained from Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Knock-Out, Functional Assay, Suspension, Staining

Journal: Advanced Science

Article Title: Schwann Cell Synthesized Cholesterol Orchestrates Peripheral Nerve Regeneration via Structural and IGF1‐Dependent Signaling Mechanisms

doi: 10.1002/advs.202520323

Figure Lengend Snippet: Squalene administration rescues the SC‐FDFT1 deficiency delayed axonal regeneration at 3dpi. (A) Immunofluorescent staining and quantifications (a1) of GAP43 + regenerating axons in the longitudinal sections of crush‐injured nerves at 3 dpi. Scale bar = 200 µm, ( n = 4). (B) Western blotting and analysis of GAP43 protein levels (b1) in the distal trunk of the injured nerves at 3 dpi ( n = 6). Data are presented as mean ± SD. Two‐way ANOVA, “ns” indicating no significance, * p < 0.05.

Article Snippet: The Fdft1 flox/flox mice, Plp CreERT2 mice and Dhh Cre mice were obtained from Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Staining, Western Blot

Journal: Advanced Science

Article Title: Schwann Cell Synthesized Cholesterol Orchestrates Peripheral Nerve Regeneration via Structural and IGF1‐Dependent Signaling Mechanisms

doi: 10.1002/advs.202520323

Figure Lengend Snippet: Squalene administration rescues the SC‐FDFT1 deficiency impaired functional regeneration at 28 dpi. (A) Representative footprints and the SFI scores (a1) at 28 dpi, ( n = 6). (B) Respective CMAP images and quantified latency (b1) and amplitudes (b2), ( n = 6). (C) Immunostaining for NF200 (red) and MBP (green) on the transverse sections of the nerve at 3 mm distal to the lesion site at 28 dpi. Scale bar = 20 µm, zoom in, 5 µm. (D) Western blotting analysis of MBP and NF200 protein levels. Quantifications shown in (d1,d2), ( n = 6). (E) TEM analysis shows the number of myelinated axons (e1), myelin thickness (e2) and G‐ratio (e3, e4) at 3 mm from the injury site at 28 dpi. Scale bar = 3 µm, zoom in, 1 µm. ( n = 3). Data are presented as mean ± SD. Two‐way ANOVA, “ns” indicating no significance, * p < 0.05.

Article Snippet: The Fdft1 flox/flox mice, Plp CreERT2 mice and Dhh Cre mice were obtained from Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Functional Assay, Immunostaining, Western Blot

Journal: Advanced Science

Article Title: Schwann Cell Synthesized Cholesterol Orchestrates Peripheral Nerve Regeneration via Structural and IGF1‐Dependent Signaling Mechanisms

doi: 10.1002/advs.202520323

Figure Lengend Snippet: FDFT1 deletion impaired SCs differentiation, is rescued by the squalene administration. (A) Schematic of the experimental design for inducing SCs differentiation with squalene or cholesterol treatments. (B) Western blot analysis of MAG protein levels, quantification shown in (b1), ( n = 4–6). (C) Immunofluorescent staining and quantification (c1) for MAG (green) and Phalloidin staining (red). Scale bar = 50 µm, ( n = 4). (D) Immunofluorescent staining and quantification (d1) for c‐Jun (red). Scale bar = 50 µm, ( n = 4). Data are presented as mean ± SD. Two‐way ANOVA, “ns” indicating no significance, * p < 0.05.

Article Snippet: The Fdft1 flox/flox mice, Plp CreERT2 mice and Dhh Cre mice were obtained from Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Western Blot, Staining

Journal: Advanced Science

Article Title: Schwann Cell Synthesized Cholesterol Orchestrates Peripheral Nerve Regeneration via Structural and IGF1‐Dependent Signaling Mechanisms

doi: 10.1002/advs.202520323

Figure Lengend Snippet: FDFT1 deletion in SCs impedes axon regeneration via squalene/cholesterol /LXRα/IGF1 signaling pathway. (A–E) Western blot analysis of the protein levels of IGF1, IGF1R, Rap1, PI3K and p‐Akt in the primary SCs of Flox, cKO and cKO+Squalene groups. (F) LXRα nuclear localization in primary cultured Flox and cKO SCs, and after squalene or cholesterol treatments, is illustrated by immunofluorescence. Scale bar = 20 µm, zoom in, 5 µm, ( n = 4). (G) Effect of LXRα agonist T0901317 on LXRα nuclear localization in cKO SCs is assessed by immunofluorescence. Scale bar = 20 µm, zoom in, 5 µm, ( n = 3). (H) IGF1 levels in the cultured supernatants of primary SCs after treating with T0901317 are measured by ELISA ( n = 3). (I) Analyze the level of LXRα nuclear import after LXR antagonist GSK2033 treatment, and the reversed effect of squalene supplementation in treated cKO SCs. Scale bar = 20 µm, zoom in, 5 µm, ( n = 3). (J) IGF1 levels in the cultured supernatants of primary SCs after treatment from (I) are measured by ELISA ( n = 3). Data are presented as mean ± SD. Two‐way ANOVA, “ns” indicating no significance, * p < 0.05.

Article Snippet: The Fdft1 flox/flox mice, Plp CreERT2 mice and Dhh Cre mice were obtained from Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Western Blot, Cell Culture, Immunofluorescence, Enzyme-linked Immunosorbent Assay

Journal: Advanced Science

Article Title: Schwann Cell Synthesized Cholesterol Orchestrates Peripheral Nerve Regeneration via Structural and IGF1‐Dependent Signaling Mechanisms

doi: 10.1002/advs.202520323

Figure Lengend Snippet: The schematic diagram illustrates the underlying mechanism of how SCs FDFT1 regulate functional peripheral nerve regeneration.

Article Snippet: The Fdft1 flox/flox mice, Plp CreERT2 mice and Dhh Cre mice were obtained from Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Functional Assay