anti scap  (Bioss)

Journal: iScience

Article Title: SESN2 maintains cartilage homeostasis by SREBP1-mediated lipid metabolism during osteoarthritis progression

doi: 10.1016/j.isci.2025.113097

Figure Lengend Snippet: SESN2 improves fatty acid metabolism disorders by inhibiting SREBP1 (A) qPCR analysis of Srebp1 in mouse primary chondrocytes pretreated with si-nc or si- Sesn2 ( n = 3). (B) Western blot analysis (left) and quantitation (right) of SREBP1 in mouse primary chondrocytes pretreated with si-nc or si- Sesn2 ( n = 3). (C) IF analysis (left) and quantitation (right) of SREBP1 in mouse primary chondrocytes pretreated with si-nc or si- Sesn2 ( n = 6). (D) SESN2 and SCAP Molecular Docking Diagrams (HDOCK Server). (E) Immunoblot analysis of mouse primary chondrocytes, followed by immunoprecipitation with anti-SESN2 and anti-SCAP beads, probed with indicated antibodies. (F) qPCR analysis of indicated genes in mouse primary chondrocytes treated as indicated for 48 h. (G and H) Western blot analysis (G) and quantitation (H) of indicated proteins in mouse primary chondrocytes treated as indicated for 48 h ( n = 3). Scale bars, 25 μm. (I) qPCR analysis of indicated genes in mouse primary chondrocytes treated as indicated for 48 h ( n = 3). (J and K) Western blot analysis (J) and quantitation (K) of mouse primary chondrocytes treated as indicated for 48 h ( n = 3). Scale bars, 50 μm. Data are expressed as mean ± SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001; NS, nonsignificant. Statistical analysis was performed by unpaired t test (A and B) and one-way ANOVA (F, H, I, and K).

Article Snippet: Pre-cleared lysates were incubated overnight at 4°C with anti-SESN2 antibody (Proteintech, #10795-1-AP, 2–4 μg) or anti-SCAP antibody (Santa Cruz, #sc-13553, 2–4 μg) or corresponding IgG control antibody.

Techniques: Western Blot, Quantitation Assay, Immunoprecipitation

Journal: Advanced Science

Article Title: A Super‐Enhancer‐Driven Transcriptional Regulatory Circuit Underlying Abiraterone Resistance in Castration‐Resistant Prostate Cancer

doi: 10.1002/advs.202501284

Figure Lengend Snippet: The super‐enhancer‐driven transcription factor loop activates the cholesterol biosynthesis. A) Reactome enrichment analysis of the genes enriched in the“Lipid biosynthetic process”of Figure . B) the RNA‐Seq data of BCL6, SMAD3, and NFIB knockdown are subjected to GSEA enrichment analysis with the gene sets associated with cholesterol homeostasis. C,D) Intracellular cholesterol, testosterone (T), and dihydrotestosterone (DHT) levels measured following the individual knockdown of BCL6, SMAD3, and NFIB in C42B‐ABi cells (C) and tumor xenografts in mice (D). One‐way ANOVA Dunnett's multiple comparisons test. n = 3. *, P <0.05; **, P < 0.01; ***, P < 0.001. E) Heatmaps illustrating the mRNA expression and binding patterns of the gene collection enriched in cholesterol homeostasis. F,G) The mRNA (F) and protein (G) expression level of indicated genes were detected in C42B‐ABi cells following silencing of BCL6, SMAD3, and NFIB. Two‐way ANOVA Dunnett's multiple comparisons test. n = 3. *, P <0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. H) The protein expression of SREBF2, HMGCR, and SCAP were measured following the over‐expression of BCL6, SMAD3, or NFIB in C42B‐ABi cells. I,J) Silencing SREBF2 by siRNA in C42B‐ABi cells (I), the cell proliferation (J) was detected. One‐way ANOVA Dunnett's multiple comparisons test. n = 3. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. K) Abiraterone dose‐response curves for C42B‐ABi cells after 3 days of SREBF2 knockdown. One‐way ANOVA Dunnett's multiple comparisons test. n = 3. *, P <0.05; **, P < 0.01; ***, P < 0.001. L) In vitro MTS proliferation assay of prostate cancer cells in the presence of different doses of fatostatin (SREBF2 inhibitor). M) Intracellular cholesterol, testosterone (T), and dihydrotestosterone (DHT) levels measured with fatostatin treatment in C42B‐ABi cells. One‐way ANOVA Dunnett's multiple comparisons test. n = 3. *, P <0.05; **, P < 0.01. N) Overexpress SREBF2 after silencing BCL6, SMAD3, and NFIB by shRNA in C42B‐ABi cells, the cell proliferation was detected. Two‐way ANOVA Dunnett's multiple comparisons test. n = 3. *, P <0.05; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: Antibody of HMGCR (#13533‐1‐AP), NFIB (#29898‐1‐AP), SMAD3 (#30130‐1‐AP), SCAP (#12266‐1‐AP), SREBF2 (#28212‐1‐AP), FDFT1 (#13128‐1‐AP), AR (#5153), PSA (#5365) were purchased from Proteintech (Rosemont, USA).

Techniques: RNA Sequencing, Knockdown, Expressing, Binding Assay, Over Expression, In Vitro, Proliferation Assay, shRNA