primary antibodies anti rap1a  (Santa Cruz Biotechnology)

Journal: Journal of Lipid Research

Article Title: Geranylgeranyl isoprenoids and hepatic Rap1a regulate basal and statin-induced expression of PCSK9

doi: 10.1016/j.jlr.2024.100515

Figure Lengend Snippet: Geranylgeranyl isoprenoid restoration suppresses statin-induced PCSK9 independent of a change in Pcsk9 mRNA and lowers LDL-C. A and B: Primary WT hepatocytes (WT HCs) were treated with vehicle, 10 μM simvastatin (simva), or simvastatin and 10 μM GGPP together for 32 h. PCSK9 protein (A) and Pcsk9 mRNA (B) levels were measured ( n = 2–4, mean ± SEM, ∗ P < 0.05, n.s., nonsignificant). C: Same as in (A) except that Rap1a −/− HCs were used and secreted PCSK9 was measured. Densitometric quantification of the immunoblot data is shown in the bar graph ( n = 4, mean ± SEM, ∗ P < 0.05, n.s., nonsignificant). D–G: Hepatic active Rap1a (D), liver PCSK9 protein (E), plasma PCSK9 (F), and plasma LDL-C (G) levels in high-fat diet-fed WT C57BL/6J male mice that were treated with vehicle, 10 mg/kg rosuvastatin (rosu), or rosu + GGOH (50 mg/kg) daily for 3 weeks. Densitometric quantification of the immunoblot data is shown in the bar graphs ( n = 4–5, mean ± SEM, ∗ P < 0.05).

Article Snippet: Rap1a activation levels were determined by Western blot using a Rap1a primary antibody (R&D Systems, catalog no.: AF3767).

Techniques: Western Blot, Clinical Proteomics

Journal: Journal of Lipid Research

Article Title: Geranylgeranyl isoprenoids and hepatic Rap1a regulate basal and statin-induced expression of PCSK9

doi: 10.1016/j.jlr.2024.100515

Figure Lengend Snippet: GGTase-I silencing and prenylation-deficient Rap1a increase PCSK9. A and B: intracellular and secreted PCSK9 protein (A) or Pcsk9 mRNA (B) from scrambled (scr) or si-Pggt1b-treated primary WT hepatocytes (WT HCs). Densitometric quantification of the immunoblot data is shown in the bar graph ( n = 4, mean ± SEM, ∗ P < 0.05, n.s., nonsignificant). C and D: Plasma PCSK9 (C) and liver Pcsk9 mRNA levels (D) from high-fat diet-fed WT C57BL/6J male mice treated with an AAV8-H1 construct encoding sh-Pggt1b or empty vector (control) ( n = 7–8, mean ± SEM, ∗ P < 0.05, n.s., nonsignificant). E–G: Rap1a −/− HCs were transfected with empty plasmid (vector) or plasmids encoding WT or prenylation-deficient Rap1a (SAAX-Rap1a), and Rap1a mRNA (E), intracellular PCSK9 (F), and Pcsk9 mRNA levels were assayed (G). Densitometric quantification of the immunoblot data is shown in the bar graph ( n = 3, mean ± SEM, ∗ P < 0.05, n.s., nonsignificant).

Article Snippet: Rap1a activation levels were determined by Western blot using a Rap1a primary antibody (R&D Systems, catalog no.: AF3767).

Techniques: Western Blot, Clinical Proteomics, Construct, Plasmid Preparation, Control, Transfection

Journal: Journal of Lipid Research

Article Title: Geranylgeranyl isoprenoids and hepatic Rap1a regulate basal and statin-induced expression of PCSK9

doi: 10.1016/j.jlr.2024.100515

Figure Lengend Snippet: Rap1a activation via 8-pCPT treatment suppresses basal and statin-induced PCSK9. A–C: WT C57BL/6J male mice that were fed with high-fat diet were treated with 1.5 mg/kg/day with 8-pCPT for 2 weeks. Hepatic GTP-Rap1a (active Rap1a) (A), plasma PCSK9 (B), and plasma total cholesterol (TC) (C) were measured. Densitometric quantification of the immunoblot data is shown in the bar graph ( n = 3–4, mean ± SEM, ∗ P < 0.05). D and E: Intracellular (D) and secreted PCSK9 (E) levels were assayed in primary WT hepatocytes (WT HCs) treated with vehicle, 10 μm simvastatin (simva), or simvastatin and 20 μm 8-pCPT together for 48 h. Densitometric quantification of the immunoblot data is shown in the bar graphs ( n = 3, mean ± SEM, ∗ P < 0.05).

Article Snippet: Rap1a activation levels were determined by Western blot using a Rap1a primary antibody (R&D Systems, catalog no.: AF3767).

Techniques: Activation Assay, Clinical Proteomics, Western Blot

Journal: Journal of Lipid Research

Article Title: Geranylgeranyl isoprenoids and hepatic Rap1a regulate basal and statin-induced expression of PCSK9

doi: 10.1016/j.jlr.2024.100515

Figure Lengend Snippet: Rap1a silencing increases PCSK9 via inducing the RhoA-ROCK pathway. A and B: RhoA levels were assayed in primary WT hepatocytes (WT HCs) treated with scrambled (scr) versus si-Rap1a (A) or in adeno-LacZ versus adeno-Rap1GAP-expressing mice liver (B). Densitometric quantification of the immunoblot data is shown in the bar graphs ( n = 4–7, mean ± SEM, ∗ P < 0.05). C and D: Scrambled (scr) or si-RhoA-treated mouse HCs were assayed for secreted and intracellular PCSK9 (C) or Pcsk9 mRNA (D). Densitometric quantification of the immunoblot data is shown in the bar graphs ( n = 4, mean ± SEM, ∗ P < 0.05). E: Mouse HCs were treated with vehicle or 1 μg/ml C3 transferase Rho inhibitor (C3), and secreted PCSK9 levels were assayed. Densitometric quantification of the immunoblot data is shown in the bar graphs ( n = 3, mean ± SEM, ∗ P < 0.05). F and G: Plasma PCSK9 (F) and plasma total cholesterol (TC) levels (G) were measured from AAV8-sh-RhoA or control AAV (Con)-treated mice fed with a high-cholesterol diet ( n = 3–4/group, mean ± SEM, ∗ P < 0.05). H and I: (H) Scramble (scr), si-Rap1a alone, or si-Rap1a + si-RhoA-treated primary HCs were assayed for secreted PCSK9. I: Same as in (H), except that scr- and si-ROCK-treated cells were used. Densitometric quantification of the immunoblot data is shown in the bar graphs ( n = 3, mean ± SEM, ∗ P < 0.05).

Article Snippet: Rap1a activation levels were determined by Western blot using a Rap1a primary antibody (R&D Systems, catalog no.: AF3767).

Techniques: Expressing, Western Blot, Clinical Proteomics, Control

Journal: Journal of Lipid Research

Article Title: Geranylgeranyl isoprenoids and hepatic Rap1a regulate basal and statin-induced expression of PCSK9

doi: 10.1016/j.jlr.2024.100515

Figure Lengend Snippet: Proposed PCSK9 regulation by statins. Statins increase PCSK9 through increasing its mRNA via SREBP2 and inhibiting its degradation by decreasing the GGPP synthesis and Rap1a activity.

Article Snippet: Rap1a activation levels were determined by Western blot using a Rap1a primary antibody (R&D Systems, catalog no.: AF3767).

Techniques: Activity Assay

Journal: Pharmaceuticals (Basel, Switzerland)

Article Title: Berberine Overcomes Gemcitabine-Associated Chemoresistance through Regulation of Rap1/PI3K-Akt Signaling in Pancreatic Ductal Adenocarcinoma.

doi: 10.3390/ph15101199

Figure Lengend Snippet: Figure 4. Rap1/PI3K-Akt signaling pathway correlates with Gemcitabine-resistance in PDAC cells. (A) Schematic of the differentially expressed gene discovery in Gem-R PDAC cells using GSE148200 and GSE140077. (B) Venn-diagram of up- and down-regulated expression (log2FC > ±1.0 and p < 0.01) of the genes in Gem-R PDAC cells. (C,D) Scatter plot of KEGG pathway enrichment analysis of up-

Article Snippet: Primary antibodies against Rap1 (1:1000, #2399; Cell Signaling Technology [CST], Danvers, MA, USA), PI3K (1:500, #4249; CST), Akt (1:1000, #4691; CST), and phospho-Akt (Ser473) (1:1000, #4060; CST) were used.

Techniques: Expressing