rap1 antibody  (Cell Signaling Technology Inc)

Journal: Diabetes, Metabolic Syndrome and Obesity

Article Title: Fu-Fang-Qi-Di-Hua-Yu-Tang Improves Diabetic Macrovascular Disease via PI3K/AKT Pathway Regulation

doi: 10.2147/DMSO.S515521

Figure Lengend Snippet: Comparison of mRNA expression via the RAP1/PI3K/AKT pathway among different groups. ( a ) Relative expression of RAP 1; ( b ) Relative expression of PI3K; ( c ) Relative expression of AKT; ( d ) Relative expression of mTOR. Compared with the control (** P< 0.01), model ( # P< 0.05, ## P< 0.01), L-FFQD ( ◆ P< 0.05, ◆◆ P< 0.01), M-FFQD ( Δ P< 0.05, ΔΔ P< 0.01), and H-FFQD ( ★ P< 0.05, ★★ P< 0.01) groups, significant differences were observed.

Article Snippet: The membrane was then incubated overnight at 4°C with primary antibodies: LC3-II/I (18725-1-AP, Proteintech, USA), NF-κB p65 (10,745-1-AP, Proteintech, USA) (1:500), RAP1 (10,840-1-AP, Proteintech, USA), PI3K (20584-1-AP, Proteintech, USA), P-PI3K (bs-6417R, Bioss, China), mTOR (#2983, CST, USA) (1:1000), P-mTOR (67778-1-Ig, Proteintech, USA) (1:2000), RAGE (16346-1-AP, Proteintech, USA), P-AKT (66444-1-Ig, Proteintech, USA) (1:3000), P62 (18,420-1-AP, Proteintech, USA), AKT (60203-2-Ig, Proteintech, USA), FOX01 (18,592-1-AP, Proteintech, USA), β-actin (66009-1-Ig, Proteintech, USA), and GAPDH (10494-1-AP, Proteintech, USA) (1:5000) antibodies.

Techniques: Comparison, Expressing, Control

Journal: Diabetes, Metabolic Syndrome and Obesity

Article Title: Fu-Fang-Qi-Di-Hua-Yu-Tang Improves Diabetic Macrovascular Disease via PI3K/AKT Pathway Regulation

doi: 10.2147/DMSO.S515521

Figure Lengend Snippet: Comparison of protein expression via the RAP1/PI3K/AKT pathway among different groups. ( a ) Electrophoresis of protein expression (A. Control group; B. Model group; C. L-FFQD group; D. M-FFQD group; E. H-FFQD group; F. ATOR-MAT group); ( b ) Relative value of RAP1; ( c ) Relative value of P-PI3K/PI3K; ( d ) Relative value of P-AKT/AKT; ( e ) Relative value of P-mTOR/mTOR; ( f ) Relative value of FOX01. Compared with the control (** P< 0.01), model ( # P< 0.05, ## P< 0.01), L-FFQD ( ◆ P< 0.05, ◆◆ P< 0.01), M-FFQD ( ΔΔ P< 0.01), and H-FFQD ( ★★ P< 0.01) groups, significant differences were observed.

Article Snippet: The membrane was then incubated overnight at 4°C with primary antibodies: LC3-II/I (18725-1-AP, Proteintech, USA), NF-κB p65 (10,745-1-AP, Proteintech, USA) (1:500), RAP1 (10,840-1-AP, Proteintech, USA), PI3K (20584-1-AP, Proteintech, USA), P-PI3K (bs-6417R, Bioss, China), mTOR (#2983, CST, USA) (1:1000), P-mTOR (67778-1-Ig, Proteintech, USA) (1:2000), RAGE (16346-1-AP, Proteintech, USA), P-AKT (66444-1-Ig, Proteintech, USA) (1:3000), P62 (18,420-1-AP, Proteintech, USA), AKT (60203-2-Ig, Proteintech, USA), FOX01 (18,592-1-AP, Proteintech, USA), β-actin (66009-1-Ig, Proteintech, USA), and GAPDH (10494-1-AP, Proteintech, USA) (1:5000) antibodies.

Techniques: Comparison, Expressing, Electrophoresis, Control

Journal: Science Advances

Article Title: Enhanced Rap1 small GTPase activity in the ventral hippocampus drives stress-induced anxiety

doi: 10.1126/sciadv.adt3163

Figure Lengend Snippet: ( A ) Experimental procedure for mice subjected to CRS. D, day. ( B and C ) Representative activity tracking and statistical analysis of time in the center and total distance traveled during OFT ( n = 12 to 13 mice per group). ( D and E ) Representative activity tracking and statistical analysis of time in open arms and open arm entries during EPMT. Same sample size as in (C). ( F ) Scheme for the analysis of total Rap1 and active Rap1. ( G ) Representative Western blots from the Con and CRS mice. ( H ) Quantification analysis of relative total Rap1 and active Rap1 expression ( n = 5 per group). ( I ) Scheme for delivering vehicle or GGTI-298 into the dHPC and vHPC. ( J ) Representative images of cannula implantation in dHPC and vHPC. ( K and L ) Representative activity tracking and statistical analysis of time in the center and total distance traveled during OFT of mice after administering vehicle and GGTI-298 in dHPC ( n = 8 to 10 mice per group). ( M and N ) Representative activity tracking and statistical analysis of time in open arms and open arm entries during EPMT. Same sample size as in (L). ( O to R ) Same as in [(K) to (N)] except that the data were from mice receiving vehicle and GGTI-298 in vHPC ( n = 9 to 11 mice per group). Data are presented as the means ± SEM. * P < 0.05 and ** P < 0.01. Two-way ANOVA [(L), (N), (P), and (R)]; Mann-Whitney U test [(C), left]; two-tailed unpaired Student’s t test [(C) right, (E), and (H)].

Article Snippet: Then, the membranes were incubated with the following primary antibodies: Kv4.2 (1:1000, Neuromab, US), phospho-Kv4.2 (1:300, Thr 607 , Santa Cruz, US), phospho-Kv4.2 (1:1000, Ser 616 , MyBioSource, US), phospho-Kv4.2 (1:1000, Thr 602 , Bioss, China), Rap1 (1:1000, Cell Signaling Technology, US), and β-actin (1:2000, Abcam, US), overnight at 4°C.

Techniques: Activity Assay, Western Blot, Expressing, MANN-WHITNEY, Two Tailed Test

Journal: Science Advances

Article Title: Enhanced Rap1 small GTPase activity in the ventral hippocampus drives stress-induced anxiety

doi: 10.1126/sciadv.adt3163

Figure Lengend Snippet: ( A ) Schematic of AAV injection to express Cre recombinase (Cre-eGFP) or eGFP alone (eGFP) under the CaMKII α promoter in the vHPC of Rap1 flx/flx mice followed by CRS and behavioral tests. ( B ) Representative images showing eGFP expression in vHPC. Scale bar, 500 μm. ( C ) Representative images of CaMKIIα immunostaining in the vCA1. Scale bar, 10 μm. ( D and E ) Representative activity tracking and quantification of time in the center and total distance traveled during OFT ( n = 14 to 15 mice per group). ( F and G ) Representative activity tracking and quantification of time in open arms and open arm entries during EPMT. Same sample size as in [(D) and (E)]. ( H ) Schematic of overexpressing Rap1-DN-mCherry or mCherry alone in the vHPC PNs followed by CRS and behavioral tests. ( I ) Representative images showing mCherry expression in vHPC. Scale bar, 500 μm. ( J ) Representative immunoblots for active Rap1 and total Rap1 and statistical analysis of the immunoblotting assay from the mCherry and Rap1-DN-mCherry mice ( n = 5 per group). ( K to N ) Same as in [(D) to (G)] except that the data were from mice infected with mCherry or Rap1-DN-mCherry ( n = 12 to 15 mice per group). ( O ) Schematic of overexpressing Rap1-CA-mCherry or mCherry alone in the vHPC PNs and subsequent behavioral tests. ( P and Q ) Same as in [(I) and (J)] except that the data were from mice infected with mCherry or Rap1-CA-mCherry ( n = 5 per group). ( R to U ) Same as in [(D) to (G)] except that the data were from mice infected with mCherry or Rap1-CA-mCherry and without subjecting to CRS ( n = 13 to 14 mice per group). Pooled data are presented as the means ± SEM. * P < 0.05, ** P < 0.01, and *** P < 0.001. Mann-Whitney U test [(E) left, (N) left, and (S) right]; two-tailed unpaired Student’s t test [(E) right, (F), (J), (L), (N) right, (Q), (S) left, and (U)].

Article Snippet: Then, the membranes were incubated with the following primary antibodies: Kv4.2 (1:1000, Neuromab, US), phospho-Kv4.2 (1:300, Thr 607 , Santa Cruz, US), phospho-Kv4.2 (1:1000, Ser 616 , MyBioSource, US), phospho-Kv4.2 (1:1000, Thr 602 , Bioss, China), Rap1 (1:1000, Cell Signaling Technology, US), and β-actin (1:2000, Abcam, US), overnight at 4°C.

Techniques: Injection, Expressing, Immunostaining, Activity Assay, Western Blot, Infection, MANN-WHITNEY, Two Tailed Test

Journal: Science Advances

Article Title: Enhanced Rap1 small GTPase activity in the ventral hippocampus drives stress-induced anxiety

doi: 10.1126/sciadv.adt3163

Figure Lengend Snippet: ( A ) Schematics of AAV injections and subsequent CRS and electrophysiological experiments. ( B ) Representative firing traces of vCA1 PNs in response to 200- and 250-pA depolarizing current injections. ( C ) Statistical analysis of AP numbers evoked by different depolarizing current injections ( n = 18 to 19 neurons from five mice per group). ( D ) Representative fAHP traces. ( E ) Statistical analysis of fAHPs. Same sample size as in (C). ( F ) Representative traces showing transient A-type K + currents obtained at various voltages in vCA1 PNs. ( G ) Statistical analysis of transient A-type K + current densities recorded at +40 mV ( n = 12 to 13 neurons from six mice per group). ( H ) Representative blots of the total Kv4.2, Kv4.2 Thr 602 , Kv4.2 Thr 607 , and Kv4.2 Ser 616 in mCherry + CRS and Rap1-DN-mCherry + CRS mice. Statistical analysis of total Kv4.2 ( I ), Kv4.2 Thr 607 ( J ), Kv4.2 Ser 616 ( K ), and Kv4.2 Thr 602 ( L ) expressions in (H) ( n = 3 mice in each group). Data are presented as the means ± SEM. * P < 0.05, ** P < 0.01, and *** p < 0.001. Two-way ANOVA with repeated measures (B); two-tailed unpaired Student’s t test [(E), (G), and (I) to (L)].

Article Snippet: Then, the membranes were incubated with the following primary antibodies: Kv4.2 (1:1000, Neuromab, US), phospho-Kv4.2 (1:300, Thr 607 , Santa Cruz, US), phospho-Kv4.2 (1:1000, Ser 616 , MyBioSource, US), phospho-Kv4.2 (1:1000, Thr 602 , Bioss, China), Rap1 (1:1000, Cell Signaling Technology, US), and β-actin (1:2000, Abcam, US), overnight at 4°C.

Techniques: Two Tailed Test

Journal: Science Advances

Article Title: Enhanced Rap1 small GTPase activity in the ventral hippocampus drives stress-induced anxiety

doi: 10.1126/sciadv.adt3163

Figure Lengend Snippet: ( A ) Experimental design for overexpressing Kv4.2-T607A (Kv4.2-T607A) or 3×FLAG alone (vehicle) in CRS mice and subsequent behavioral tests. ( B ) Representative blots and comparisons of the Kv4.2 Thr 607 expression from vehicle and Kv4.2-T607A mice ( n = 3 mice per group). ( C ) Representative traces showing transient A-type K + currents obtained at various voltages in vCA1 PNs and comparisons of transient A-type K + current densities recorded at +40 mV ( n = 7 to 8 neurons from three mice per group). ( D ) Representative activity tracking in OFT. ( E ) Statistical analysis of time in the center and total distance traveled during OFT ( n = 13 to 15 mice per group). ( F ) Representative activity tracking in EPMT. ( G ) Statistical analysis of time in open arms and open arm entries during EPMT. Same sample size as in (E). ( H ) Representative firing traces of vCA1 PNs in response to depolarizing current injection. ( I ) Statistical analysis of AP numbers evoked by different depolarizing current injections in (H) ( n = 15 to 19 neurons from four to five mice per group). ( J ) Representative fAHP traces. ( K ) Statistical analysis of fAHPs in (J). Same sample size as in (I). ( L ) Schematic depicting the virus injection strategy for inhibiting phosphorylation of Kv4.2 at Thr 607 in Rap1-CA mice and subsequent behavioral tests. ( M to T ) Same as in [(D) to (K)] except that the data were from vehicle + Rap1-CA and Kv4.2-T607A + Rap1-CA mice [(M) to (P), n = 10 to 14 mice per group; (Q) to (T), n = 13 to 15 neurons from four mice per group]. Data are presented as the means ± SEM. * P < 0.05, ** P < 0.01, and *** P < 0.001. Two-way ANOVA with repeated measures [(I) and (R)]; Mann-Whitney U test [(N) left, and (P) left]; two-tailed unpaired Student’s t test [(B), (C), (E), (G), (K), (N) right, (P) right, and (T)].

Article Snippet: Then, the membranes were incubated with the following primary antibodies: Kv4.2 (1:1000, Neuromab, US), phospho-Kv4.2 (1:300, Thr 607 , Santa Cruz, US), phospho-Kv4.2 (1:1000, Ser 616 , MyBioSource, US), phospho-Kv4.2 (1:1000, Thr 602 , Bioss, China), Rap1 (1:1000, Cell Signaling Technology, US), and β-actin (1:2000, Abcam, US), overnight at 4°C.

Techniques: Expressing, Activity Assay, Injection, Virus, Phospho-proteomics, MANN-WHITNEY, Two Tailed Test

Journal: Science Advances

Article Title: Enhanced Rap1 small GTPase activity in the ventral hippocampus drives stress-induced anxiety

doi: 10.1126/sciadv.adt3163

Figure Lengend Snippet: Chronic stress triggers prolonged Rap1 activation in vHPC PNs, facilitating the phosphorylation of Kv4.2 at Thr 607 and diminishing its channel activity. Consequently, when exposed to external anxiogenic environmental cues, the responsiveness of these neurons is heightened, leading to an anxiogenic effect.

Article Snippet: Then, the membranes were incubated with the following primary antibodies: Kv4.2 (1:1000, Neuromab, US), phospho-Kv4.2 (1:300, Thr 607 , Santa Cruz, US), phospho-Kv4.2 (1:1000, Ser 616 , MyBioSource, US), phospho-Kv4.2 (1:1000, Thr 602 , Bioss, China), Rap1 (1:1000, Cell Signaling Technology, US), and β-actin (1:2000, Abcam, US), overnight at 4°C.

Techniques: Activation Assay, Phospho-proteomics, Activity Assay

Journal: Biochemical and biophysical research communications

Article Title: FAK regulates trophoblast functions of invasion and proliferation through Rap1 pathway in early-onset preeclampsia.

doi: 10.1016/j.bbrc.2025.151788

Figure Lengend Snippet: Fig. 4. FAK regulates the function of trophoblasts via the Rap1 pathway. A: KEGG analysis of Y15-treated HTR-8/SVneo cells. B: Western blotting of Rap1 in HTR8 cells treated with Y15. C: Western blotting of Rap1 in normal and EOPE placentas. A representative image is shown. D: Rap1 immunostaining of placenta from full- term normotensive pregnancies and EOPE patients. Bar = 100 μm. All statistical data were analyzed by Student’s t-test. Data are shown as mean ± SD. *P < 0.05.

Article Snippet: Primary antibodies were used to investigate these proteins: anti-FAK (1:1000, Wanleibio, China), anti-pY397FAK (1:1000, Cell Signaling Technology, USA), anti-PCNA (1:1000, Proteintech, USA), anti-Rap1(1:1000, Proteintech, USA), anti-N-cadherin (1:1000, CST, USA), and anti-β-actin (1:1000, Proteintech, USA).

Techniques: Western Blot, Immunostaining

Journal: Biochemical and biophysical research communications

Article Title: FAK regulates trophoblast functions of invasion and proliferation through Rap1 pathway in early-onset preeclampsia.

doi: 10.1016/j.bbrc.2025.151788

Figure Lengend Snippet: Fig. 5. FAK is downregulated in PE-like mouse models. A: Systolic blood pressure in L-NAME group and control group was monitored at GD 6.5, 8.5, 10.5, 12.5, 14.5, 16.5, 17.5. L-NAME: N-nitro-L-Arginine Methyl Ester. B: H&E staining of mice placentas at GD 17.5. The ratio of the placenta labyrinth zone (Lz) area to the junctional zone (Jz) area in the control group (n = 4) and L-NAME group (n = 4) was statistically calculated. Bar = 500 μm. C: HE staining of mouse kidney showed glomerular destruction in L-NAME mice (n = 3 from 3 different dams). Bar = 100 μm. D: Changes in urinary protein levels in in L- NAME (n = 5) and control (n = 5) group mouse. E: Immunofluorescence staining of FAK, pY397FAK and Rap1 in the placenta of mice at GD 17.5. Bar = 500 μm (a–c), Bar = 200 μm (d). F and G: Western blot analysis of FAK, pY397FAK and Rap1 expression in L- NAME (placenta n = 7 from 7 dams) and control (placenta n = 7 from 7 dams) group mouse placentas on GD 17.5. Statistical data were analyzed by Student’s t-test. Data are shown as mean ± SD. *P < 0.05. Dc: decidua. L-NAME: N-nitro-L-Arginine Methyl Ester.

Article Snippet: Primary antibodies were used to investigate these proteins: anti-FAK (1:1000, Wanleibio, China), anti-pY397FAK (1:1000, Cell Signaling Technology, USA), anti-PCNA (1:1000, Proteintech, USA), anti-Rap1(1:1000, Proteintech, USA), anti-N-cadherin (1:1000, CST, USA), and anti-β-actin (1:1000, Proteintech, USA).

Techniques: Control, Staining, Immunofluorescence, Western Blot, Expressing