Journal: Cancer Science

Article Title: Squalene synthase predicts poor prognosis in stage I‐III colon adenocarcinoma and synergizes squalene epoxidase to promote tumor progression

doi: 10.1111/cas.15248

Figure Lengend Snippet: Squalene synthase (FDFT1) was upregulated in stage I‐III colon adenocarcinoma (COAD) tissues and colon cancer cell lines. A, Comparison of FDFT1 expression between 217 COAD and paired adjacent normal (AN) tissues from tissue microarray (TMA). B, Comparison of FDFT1 expression in different TNM stages (I‐III) from TMA. C, Comparison of FDFT1 expression in different T stages (1‐3, 4) from TMA. D, Comparison of FDFT1 expression in different differentiation grades (well/moderate, poor/mucinous) from TMA. Mean ± SD. E, FDFT1 expression in two paired AN and COAD tissues were detected by immunohistochemical analysis. FDFT1 low and FDFT1 high in COAD tissues are shown below. Scale bars, 50 μm. F, FDFT1 mRNA level in normal colon mucosal epithelial cells (NCM460) and colon cancer cell lines. NCM460 was used as control, housekeeping gene ACTB was used loading control. * P < .05, *** P < .001

Article Snippet: Membranes were incubated with primary Abs against FDFT1 (1:1000 dilution; Cat# 13128‐1‐AP; Proteintech), NAT8 (1:500 dilution; Cat# A7759; Abclonal), SQLE (1:1000 dilution; Cat# 12544‐1‐AP; Proteintech), Tubulin (1:5000 dilution; Cat# E7; DSHB), and β‐actin (1:5000 dilution; Cat# AC026; Abclonal) after being blocked at 4°C.

Techniques: Comparison, Expressing, Microarray, Immunohistochemical staining, Control

Journal: Cancer Science

Article Title: Squalene synthase predicts poor prognosis in stage I‐III colon adenocarcinoma and synergizes squalene epoxidase to promote tumor progression

doi: 10.1111/cas.15248

Figure Lengend Snippet: Comparison of clinicopathologic features of patients with stage I‐III colon adenocarcinoma between low and high expression of squalene synthase (FDFT1)

Article Snippet: Membranes were incubated with primary Abs against FDFT1 (1:1000 dilution; Cat# 13128‐1‐AP; Proteintech), NAT8 (1:500 dilution; Cat# A7759; Abclonal), SQLE (1:1000 dilution; Cat# 12544‐1‐AP; Proteintech), Tubulin (1:5000 dilution; Cat# E7; DSHB), and β‐actin (1:5000 dilution; Cat# AC026; Abclonal) after being blocked at 4°C.

Techniques: Comparison, Expressing

Journal: Cancer Science

Article Title: Squalene synthase predicts poor prognosis in stage I‐III colon adenocarcinoma and synergizes squalene epoxidase to promote tumor progression

doi: 10.1111/cas.15248

Figure Lengend Snippet: Univariate and multivariate analyses for overall survival (OS) and relapse‐free survival (RFS) in patients with stage I‐III colon adenocarcinoma

Article Snippet: Membranes were incubated with primary Abs against FDFT1 (1:1000 dilution; Cat# 13128‐1‐AP; Proteintech), NAT8 (1:500 dilution; Cat# A7759; Abclonal), SQLE (1:1000 dilution; Cat# 12544‐1‐AP; Proteintech), Tubulin (1:5000 dilution; Cat# E7; DSHB), and β‐actin (1:5000 dilution; Cat# AC026; Abclonal) after being blocked at 4°C.

Techniques: Expressing

Journal: Cancer Science

Article Title: Squalene synthase predicts poor prognosis in stage I‐III colon adenocarcinoma and synergizes squalene epoxidase to promote tumor progression

doi: 10.1111/cas.15248

Figure Lengend Snippet: Upregulation of squalene synthase (FDFT1) was associated with poor prognosis in stage I‐III colon adenocarcinoma (COAD). A, Kaplan‐Meier curve for overall survival (OS) according to FDFT1 expression in 233 cases of stage I‐III COAD. FDFT1 high expression indicated a lower OS rate (log‐rank P < .001). B, C, Kaplan‐Meier curves for OS according to FDFT1 expression in 115 and 118 cases of right‐ and left‐sided COAD, respectively. FDFT1 high expression indicated a lower OS rate (both log‐rank P < .001). D, Nomogram for predicting 5‐year OS of stage I‐III COAD. CEA, carcinoembryonic antigen. E, Receiver operating characteristic (ROC) analysis of the nomogram for predicting 5‐year OS (area under the ROC curve [AUC] = 0.871). F, Kaplan‐Meier curve for relapse‐free survival (RFS) according to FDFT1 expression in 233 cases of stage I‐III COAD. FDFT1 high expression indicated a lower RFS rate (log‐rank P < .001). G, H, Kaplan‐Meier curves for RFS according to FDFT1 expression in 115 and 118 cases of right‐ and left‐sided COAD, respectively. FDFT1 high expression indicated a lower RFS rate (log‐rank P < .001 and P =.001, respectively). I, Nomogram for predicting 5‐year RFS of stage I‐III COAD. J, ROC analysis of the nomogram for predicting 5‐year RFS (AUC = 0.863)

Article Snippet: Membranes were incubated with primary Abs against FDFT1 (1:1000 dilution; Cat# 13128‐1‐AP; Proteintech), NAT8 (1:500 dilution; Cat# A7759; Abclonal), SQLE (1:1000 dilution; Cat# 12544‐1‐AP; Proteintech), Tubulin (1:5000 dilution; Cat# E7; DSHB), and β‐actin (1:5000 dilution; Cat# AC026; Abclonal) after being blocked at 4°C.

Techniques: Expressing

Journal: Cancer Science

Article Title: Squalene synthase predicts poor prognosis in stage I‐III colon adenocarcinoma and synergizes squalene epoxidase to promote tumor progression

doi: 10.1111/cas.15248

Figure Lengend Snippet: Squalene synthase (FDFT1) deficiency attenuated proliferation of HCT116 and HT29 colon cancer cells. A, B, After knockdown (KD) of FDFT1, the protein level of FDFT1 in HCT116 or HT29 cells was detected by western blot analysis; shcontrol (SC) as control group. C, D, After KD of FDFT1, the proliferation of HCT116 or HT29 cells was measured by counting cell numbers. E, After KD of FDFT1, the proliferation of HCT116 or HT29 cells was measured by EdU staining. F, Statistical results of (E). G, Colony formation of HCT116 or HT29 cells was examined by crystal violet staining after KD of FDFT1. H, Statistical results of (G). I, J, After separate overexpression (OE) of FDFT1 in HCT116 or HT29 cells, the protein level of FDFT1 was detected by western blot analysis; vector as control group. K, Colony formation of HCT116 cells or HT29 cells was examined by crystal violet staining after OE of FDFT1. L, Statistical results of (K). Mean ± SEM. * P < .05; ** P < .01; *** P < .001; ns, no significance

Article Snippet: Membranes were incubated with primary Abs against FDFT1 (1:1000 dilution; Cat# 13128‐1‐AP; Proteintech), NAT8 (1:500 dilution; Cat# A7759; Abclonal), SQLE (1:1000 dilution; Cat# 12544‐1‐AP; Proteintech), Tubulin (1:5000 dilution; Cat# E7; DSHB), and β‐actin (1:5000 dilution; Cat# AC026; Abclonal) after being blocked at 4°C.

Techniques: Knockdown, Western Blot, Control, Staining, Over Expression, Plasmid Preparation

Journal: Cancer Science

Article Title: Squalene synthase predicts poor prognosis in stage I‐III colon adenocarcinoma and synergizes squalene epoxidase to promote tumor progression

doi: 10.1111/cas.15248

Figure Lengend Snippet: Knockdown (KD) of squalene synthase (FDFT1) induced accumulation of N ‐acetyltransferase 8 (NAT8) and D‐pantethine to reduce reactive oxygen species level and inhibit proliferation of HT29 cells. A, Volcano plots of RNA sequencing analysis between shControl and FDFT1 KD HT29 cells; 1940 genes were upregulated, and 1828 genes were downregulated. Arrow indicates NAT8. B, Volcano plots of untargeted metabolomics analysis between shControl and FDFT1 KD HT29 cells. Arrow indicates D‐pantethine. C, Fragments per kilobase of transcript per million mapped reads (FPKM) value of NAT8 in shControl and FDFT1 KD HT29 cells from (A). D, Value of D‐pantethine in shControl and FDFT1 KD HT29 cells from (B). E, Schematic diagram of FDFT1 regulating NAT8 and D‐pantethine. Red boxes represent genes; blue boxes represent metabolites. Downregulation of FDFT1 (green) leads to upregulation of NAT8 and D‐pantethine (red). F, shControl and FDFT1 KD HT29 cells were pretreated with 1 mmol L −1 N ‐acetyl‐l‐cysteine NAC for 1 h, then analyzed by flow cytometry through DCFH‐DA staining. G, NAT8 was overexpressed in shControl and FDFT1 KD HT29 cells. Protein levels of FDFT1 and NAT8 (red star) were detected by western blot analysis; β‐actin as a loading control. Green star indicates degradation of NAT8. H‐J, Cell proliferation and colony formation of these cells were measured by counting cell numbers and crystal violet staining. K, L, shControl and FDFT1 KD HT29 cells were treated with D‐pantethine at concentrations of 0, 5, 10 μmol L −1 , respectively, then stained by crystal violet. Mean ± SEM. * P < .05; ** P < .01; *** P < .001; ns, no significance

Article Snippet: Membranes were incubated with primary Abs against FDFT1 (1:1000 dilution; Cat# 13128‐1‐AP; Proteintech), NAT8 (1:500 dilution; Cat# A7759; Abclonal), SQLE (1:1000 dilution; Cat# 12544‐1‐AP; Proteintech), Tubulin (1:5000 dilution; Cat# E7; DSHB), and β‐actin (1:5000 dilution; Cat# AC026; Abclonal) after being blocked at 4°C.

Techniques: Knockdown, RNA Sequencing, Flow Cytometry, Staining, Western Blot, Control

Journal: Cancer Science

Article Title: Squalene synthase predicts poor prognosis in stage I‐III colon adenocarcinoma and synergizes squalene epoxidase to promote tumor progression

doi: 10.1111/cas.15248

Figure Lengend Snippet: Combined inhibition of squalene synthase (FDFT1) and squalene epoxidase (SQLE) suppressed growth of colon cancer cells and xenograft tumors. A, After separate or double knockdown (KD) of FDFT1 and SQLE, the protein levels of FDFT1 and SQLE in HT29 cells were detected by western blot analysis; shControl, control group. B, Colony formation of cells in (A) were measured by crystal violet staining. C, Statistical results of (B). D, Intracellular total cholesterol level of shControl, FDFT1 KD, SQLE KD, and FDFT1/SQLE HT29 cells were measured. E, HT29 cells were treated with lapaquistat (La) and terbinafine (Te) at different concentrations for 48 or 72 h, then stained with EdU. F, After combined treatment with La and Te, the colony formation of HCT116 and HT29 cells was measured by crystal violet staining. G, Statistical results of (F). H, Tumor growth of shControl, FDFT1 KD, SQLE KD, and FDFT1/SQLE KD HT29 cells in nude mice after injection for 1 wk. I, J, Images and weights of the tumors in (H) removed from nude mice. K, Immunohistochemical analysis of Ki‐67 in xenograft tumors from (I). Scale bars, 50 μm. L, Statistical results of (K). M, FDFT1 and N ‐acetyltransferase 8 (NAT8) mRNA levels in tumors derived from shControl and FDFT1 KD HT29 cells. Mean ± SEM. * P < .05, ** P < .01, *** P < .001

Article Snippet: Membranes were incubated with primary Abs against FDFT1 (1:1000 dilution; Cat# 13128‐1‐AP; Proteintech), NAT8 (1:500 dilution; Cat# A7759; Abclonal), SQLE (1:1000 dilution; Cat# 12544‐1‐AP; Proteintech), Tubulin (1:5000 dilution; Cat# E7; DSHB), and β‐actin (1:5000 dilution; Cat# AC026; Abclonal) after being blocked at 4°C.

Techniques: Inhibition, Knockdown, Western Blot, Control, Staining, Injection, Immunohistochemical staining, Derivative Assay

Journal: Cancer Science

Article Title: Squalene synthase predicts poor prognosis in stage I‐III colon adenocarcinoma and synergizes squalene epoxidase to promote tumor progression

doi: 10.1111/cas.15248

Figure Lengend Snippet: Schematic diagram showing synergistic function of squalene synthase (FDFT1) and squalene epoxidase (SQLE) in promoting colon adenocarcinoma (COAD). FDFT1 induces accumulation of reactive oxygen species (ROS) level and promotes colon cancer cell proliferation through upregulation of D‐pantethine and N ‐acetyltransferase 8 (NAT8). FDFT1 also synergizes SQLE to facilitate COAD progression

Article Snippet: Membranes were incubated with primary Abs against FDFT1 (1:1000 dilution; Cat# 13128‐1‐AP; Proteintech), NAT8 (1:500 dilution; Cat# A7759; Abclonal), SQLE (1:1000 dilution; Cat# 12544‐1‐AP; Proteintech), Tubulin (1:5000 dilution; Cat# E7; DSHB), and β‐actin (1:5000 dilution; Cat# AC026; Abclonal) after being blocked at 4°C.

Techniques:

Journal: The Journal of Immunology Author Choice

Article Title: Mevalonate Biosynthesis Intermediates Are Key Regulators of Innate Immunity in Bovine Endometritis

doi: 10.4049/jimmunol.1501080

Figure Lengend Snippet: Cholesterol biosynthesis pathway. Cholesterol is the predominant sterol in vertebrates, and in eukaryotes, the mevalonate pathway is the main synthesis pathway for cholesterol. Acetyl CoA and acetoacetyl CoA are converted via the isoprenoids (e.g., FPP and GGPP) to squalene. Three of the key enzymes in this pathway are HMGCR, FDPS, and FDFT1. The importance of cholesterol to a variety of cellular processes means that cholesterol concentration within the cell is tightly regulated. Consequently, cholesterol synthesis is closely linked to cholesterol uptake via receptors such as the low-density lipoprotein receptor (LDLR), and export from the cell via transporters such as the ATP-binding cassette transporter A1 (ABCA1).

Article Snippet: Membranes were probed with Abs targeting FDFT1 (NBP1-54855; Novus Biologicals, Cambridge, U.K.), and HMGCR (ab98018; Abcam, Cambridge, U.K.), using Abs selected based on recognition of immunoreactive proteins of appropriate m.w.

Techniques: Concentration Assay, Binding Assay

Journal: The Journal of Immunology Author Choice

Article Title: Mevalonate Biosynthesis Intermediates Are Key Regulators of Innate Immunity in Bovine Endometritis

doi: 10.4049/jimmunol.1501080

Figure Lengend Snippet: Endometrial cell inflammatory responses to LPS are attenuated by blocking FDFT1, but not FDPS or HMGCR. Endometrial epithelial ( A , B , E , F , I , and J ) and stromal ( C , D , G , H , K , and L ) cells were treated with control medium of medium containing atorvastatin (0.1–10 μM), etidronate (1–100 μM), squalestatin (0.1–10 μM), or 25 nM dexamethasone for 24 h. Cells were subsequently challenged with control medium or 100 ng/ml LPS for a further 24 h. Supernatants were collected for analysis of IL-6 (A, C, E, G, I, and K) and CXCL8 (B, D, F, H, J, and L) by ELISA (bars), and cells were lysed and stored in RIPA buffer for analysis of cholesterol (CHO) concentration by enzymatic assay (red squares). Data are presented as mean (SEM) from four independent experiments. Data were analyzed by ANOVA and Dunnett’s pairwise multiple comparison t test; values differ from LPS (0 + LPS), * p < 0.05, ** p < 0.01, *** p < 0.001; or values differ from control (0), * p < 0.05, ** p < 0.01. ND, not detected.

Article Snippet: Membranes were probed with Abs targeting FDFT1 (NBP1-54855; Novus Biologicals, Cambridge, U.K.), and HMGCR (ab98018; Abcam, Cambridge, U.K.), using Abs selected based on recognition of immunoreactive proteins of appropriate m.w.

Techniques: Blocking Assay, Control, Enzyme-linked Immunosorbent Assay, Concentration Assay, Enzymatic Assay, Comparison

Journal: The Journal of Immunology Author Choice

Article Title: Mevalonate Biosynthesis Intermediates Are Key Regulators of Innate Immunity in Bovine Endometritis

doi: 10.4049/jimmunol.1501080

Figure Lengend Snippet: Endometrial cell inflammatory responses to LPS are attenuated in a dose-dependent manner by inhibition of FDFT1. Endometrial epithelial ( A , B , E , and F ) and stromal ( C , D , G , and H ) cells were treated with control medium or medium containing CP-34086894 [(A–D) 0.01–100 μM], squalestatin [(E–H) 0.01–100 μM], or dexamethasone (Dex; 25 nM) for 24 h. After treatment, cells were challenged with control medium or 100 ng/ml LPS for 24 h. Supernatants were collected for analysis of IL-6 (A, E, C, and G) and CXCL8 (B, F, D, and H) by ELISA. Data are presented as mean (SEM) from four independent experiments. Data were analyzed by ANOVA and Dunnett’s pairwise multiple comparison t test; values differ from LPS treatment, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Membranes were probed with Abs targeting FDFT1 (NBP1-54855; Novus Biologicals, Cambridge, U.K.), and HMGCR (ab98018; Abcam, Cambridge, U.K.), using Abs selected based on recognition of immunoreactive proteins of appropriate m.w.

Techniques: Inhibition, Control, Enzyme-linked Immunosorbent Assay, Comparison

Journal: The Journal of Immunology Author Choice

Article Title: Mevalonate Biosynthesis Intermediates Are Key Regulators of Innate Immunity in Bovine Endometritis

doi: 10.4049/jimmunol.1501080

Figure Lengend Snippet: RNA interference of HMGCR or FDFT1 modulates endometrial stromal cell innate immune responses to LPS. Endometrial stromal cells were transfected with scrambled siRNA (siScram) or siRNA targeting HMGCR (siHMG) or FDFT1 (siFDFT1) for 48 h. After transfection, cells were challenged with control medium or 100 ng/ml LPS for 24 h. Supernatants were collected for analysis of IL-6 ( A ) and CXCL8 ( B ) by ELISA. Data are presented as mean (SEM) from four independent experiments. Data were analyzed by ANOVA and Dunnett’s pairwise multiple comparison t test; values differ from LPS treatment, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Membranes were probed with Abs targeting FDFT1 (NBP1-54855; Novus Biologicals, Cambridge, U.K.), and HMGCR (ab98018; Abcam, Cambridge, U.K.), using Abs selected based on recognition of immunoreactive proteins of appropriate m.w.

Techniques: Transfection, Control, Enzyme-linked Immunosorbent Assay, Comparison

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