rabbit anti rab8b  (Proteintech)

Journal: JACC: Basic to Translational Science

Article Title: Myocardial Infarction Suppresses Protein Synthesis and Causes Decoupling of Transcription and Translation

doi: 10.1016/j.jacbts.2024.02.014

Figure Lengend Snippet: Validation of RAB8B Expression in the Heart After the Treatment of MI (A, B) Spatial transcriptomics analysis of Rab8b and Zc3hav1 expression at different time points as indicated, both are MI-induced genes. (C) Immunofluorescence staining of RAB8B in hearts after MI. Arrowheads indicate CMs with high RAB8B expression in the border zone. Scale bar = 25 μm. (D) Quantification of RAB8B expression in CMs localize at border zone or remote zone. 80 CMs in each group from 4 biological mouse hearts. Data are presented as mean ± SEM and were compared by means of the Wilcoxon signed-rank test. ∗∗∗ P < 0.001. BZ = border zone; CM = cardiomyocyte; DAPI = 6-diamino-2-phenylindole; IZ = ischemic zone; MI = myocardial infarction; RZ = remote zone; WGA = wheat germ agglutinin.

Article Snippet: The following antibodies were used: rabbit anti-Rab8b (ab222017, Abcam) and rabbit anti-Zc3hav1 (ab154680, Abcam).

Techniques: Expressing, Immunofluorescence, Staining

Journal: JACC: Basic to Translational Science

Article Title: Myocardial Infarction Suppresses Protein Synthesis and Causes Decoupling of Transcription and Translation

doi: 10.1016/j.jacbts.2024.02.014

Figure Lengend Snippet: Validation of RAB8B Expression in the Heart After the Treatment of MI (A, B) Spatial transcriptomics analysis of Rab8b and Zc3hav1 expression at different time points as indicated, both are MI-induced genes. (C) Immunofluorescence staining of RAB8B in hearts after MI. Arrowheads indicate CMs with high RAB8B expression in the border zone. Scale bar = 25 μm. (D) Quantification of RAB8B expression in CMs localize at border zone or remote zone. 80 CMs in each group from 4 biological mouse hearts. Data are presented as mean ± SEM and were compared by means of the Wilcoxon signed-rank test. ∗∗∗ P < 0.001. BZ = border zone; CM = cardiomyocyte; DAPI = 6-diamino-2-phenylindole; IZ = ischemic zone; MI = myocardial infarction; RZ = remote zone; WGA = wheat germ agglutinin.

Article Snippet: Figure 5 Validation of RAB8B Expression in the Heart After the Treatment of MI (A, B) Spatial transcriptomics analysis of Rab8b and Zc3hav1 expression at different time points as indicated, both are MI-induced genes. (C) Immunofluorescence staining of RAB8B in hearts after MI.

Techniques: Biomarker Discovery, Expressing, Immunofluorescence, Staining

Journal: European Journal of Immunology

Article Title: ACKR1 favors transcellular over paracellular T‐cell diapedesis across the blood‐brain barrier in neuroinflammation in vitro

doi: 10.1002/eji.202149238

Figure Lengend Snippet: Candidate genes selected from RNAseq

Article Snippet: RAB8B , Proteintech , 11792‐1‐AP , Polyclonal , Rabbit , Mouse, Human, Dog , IgG , 1/100 , Dotty perinuclear and cytoplasmatic staining.

Techniques:

Journal: European Journal of Immunology

Article Title: ACKR1 favors transcellular over paracellular T‐cell diapedesis across the blood‐brain barrier in neuroinflammation in vitro

doi: 10.1002/eji.202149238

Figure Lengend Snippet: Reagents used for immunofluorescence staining of candidate genes

Article Snippet: RAB8B , Proteintech , 11792‐1‐AP , Polyclonal , Rabbit , Mouse, Human, Dog , IgG , 1/100 , Dotty perinuclear and cytoplasmatic staining.

Techniques: Immunofluorescence, Staining

Journal: The Journal of Biological Chemistry

Article Title: Rab8 attenuates Wnt signaling and is required for mesenchymal differentiation into adipocytes

doi: 10.1016/j.jbc.2021.100488

Figure Lengend Snippet: Loss of Rab8a led to an elevated pLrp6 and an altered vesicular Lrp6 compartment. A , western blot showed Rab8b knockdown efficiency in Rab8b KD and Rab8a −/− ;Rab8b KD MEFs. Note Rab8a and Rab8b are similar in size but could be distinguished as two isoform-specific bands. B , western blot using lysates of MEFs after 16 passages showed continued Rab8b KD. β-Actin was used as a loading control. C , serum-starved WT, Rab8a −/− , Rab8b KD , and Rab8a −/− ;Rab8b KD MEFs were transiently transfected with 0.5-μg pCS2–LRP6 GFP for 16 h. Cells were then stimulated by vehicle (Dulbecco's modified Eagle's medium) or Wnt3a for 15 min. Indirect immunofluorescence for GFP was performed to locate LRP6. Note that LRP6 puncta were rare in vehicle-treated WT MEFs but became prominent in Wnt3a-stimulated cells. D , the numbers of LRP6 puncta were manually counted from individual cells for each condition. Experiments were repeated five times. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001 when compared with vehicle-treated WT. E – G , sucrose density cell fractionation assays were performed on serum-starved WT, Rab8a −/− , or Rab8b KD MEFs stimulated with Wnt3a or vehicle. Western blots for total Lpr6 or pLrp6 were performed. Longer exposure of pLrp6 blot for WT in panel E showed minimal but detectable signal in unstimulated condition. These data represent at least three independent experiments. H and I , serum-starved WT, Rab8a −/− , Rab8b KD , and Rab8a −/− ;Rab8b KD MEFs were transiently transfected with pCS2–LRP6 GFP for 24 h. Cells were then stained for LRP6-GFP and phalloidin. The numbers of peripheral LRP6 puncta ( arrows ), based on phalloidin staining, were manually counted for individual cells of designated genotypes. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001, when compared with WT. MEFs, mouse embryonic fibroblasts.

Article Snippet: Rab8b-specific lentiviral shRNA construct targeting against the 3'UTR of mouse Rab8b was constructed by inserting the annealed complementary oligonucleotides (5'-CCGGGCCAAGAACTAACAGAACTTTCCATGGAAAGTTCTGTTAGTTCTTGGCTTTTTG-3' and 5'-AATTCAAAAAGCCAAGAACTAACAGAACTTTCCATGGAAAGTTCTGTTAGTTCTTGGC-3') into pLK0.1 lentiviral vector (Addgene) between AgeI and EcoRI sites.

Techniques: Western Blot, Transfection, Modification, Immunofluorescence, Cell Fractionation, Staining

Journal: The Journal of Biological Chemistry

Article Title: Rab8 attenuates Wnt signaling and is required for mesenchymal differentiation into adipocytes

doi: 10.1016/j.jbc.2021.100488

Figure Lengend Snippet: Combined loss of Rab8a and Rab8b diminishes lipid-droplet formation ex vivo . A – E , representative immunofluorescence micrographs of lipid-droplet staining (BODIPY 493/503; green ) in WT, Rab8a −/− , Rab8b KD , Rab8a −/− ;Rab8b KD , or Kif3a −/− MEFs after 2 weeks of induced adipocyte differentiation. Nuclei were stained in blue . F , quantifications for individual lipid-droplet diameter. G , quantifications for the overall areas of lipid droplets in WT, Rab8a −/− , Rab8b KD , Rab8a −/− ;Rab8b KD , or Kif3a −/− cultures. Data in panels F and G represent 171, 110, 242, 101, and 101 lipid droplets for each of above genotypes. p values were determined by one-way ANOVA. H , quantitative real-time RT-PCR of proadipocyte marker transcripts Glut4, Fabp4, and PPAR-γ in differentiated WT or Rab8a −/− ;Rab8b KD MEFs. p values were determined by Student's t test. ∗ p < 0.05, when compared with WT. MEFs, mouse embryonic fibroblasts.

Article Snippet: Rab8b-specific lentiviral shRNA construct targeting against the 3'UTR of mouse Rab8b was constructed by inserting the annealed complementary oligonucleotides (5'-CCGGGCCAAGAACTAACAGAACTTTCCATGGAAAGTTCTGTTAGTTCTTGGCTTTTTG-3' and 5'-AATTCAAAAAGCCAAGAACTAACAGAACTTTCCATGGAAAGTTCTGTTAGTTCTTGGC-3') into pLK0.1 lentiviral vector (Addgene) between AgeI and EcoRI sites.

Techniques: Ex Vivo, Immunofluorescence, Staining, Quantitative RT-PCR, Marker

Journal: The Journal of Biological Chemistry

Article Title: Rab8 attenuates Wnt signaling and is required for mesenchymal differentiation into adipocytes

doi: 10.1016/j.jbc.2021.100488

Figure Lengend Snippet: Intact cilia growth and partial structural defects in Rab8-deficient MEFs. A , representative immunofluorescent images of the primary cilia that were labeled by acetylated tubulin ( green ). The basal body was marked by γ-tubulin ( red ) and nuclei by DAPI ( blue ) in WT, Rab8a −/− , Rab8b KD , or Rab8a −/− ;Rab8b KD , and Kif3a −/−−/− MEFs. Cilia induction was observed after growth arrest on a time course of 48 h after serum starvation. B and C , quantification of the percentage of ciliated cells and the average length of a cilium that was measured in pixels. Graphs showed quantifications from six fields of each genotype, containing approximately 20 to 30 cells per time point. Experiments were repeated five times. D , representative immunofluorescence of ciliary defects. White arrowheads pointed to multiciliated cells, multiple cilia at one basal body, or multiple basal bodies in Rab8a −/− or Rab8a −/− ;Rab8b KD cells. MEFs, mouse embryonic fibroblasts.

Article Snippet: Rab8b-specific lentiviral shRNA construct targeting against the 3'UTR of mouse Rab8b was constructed by inserting the annealed complementary oligonucleotides (5'-CCGGGCCAAGAACTAACAGAACTTTCCATGGAAAGTTCTGTTAGTTCTTGGCTTTTTG-3' and 5'-AATTCAAAAAGCCAAGAACTAACAGAACTTTCCATGGAAAGTTCTGTTAGTTCTTGGC-3') into pLK0.1 lentiviral vector (Addgene) between AgeI and EcoRI sites.

Techniques: Labeling, Immunofluorescence

Journal: The Journal of Biological Chemistry

Article Title: Rab8 attenuates Wnt signaling and is required for mesenchymal differentiation into adipocytes

doi: 10.1016/j.jbc.2021.100488

Figure Lengend Snippet: Aberrant Fzd2 translocation and processing in Rab8-deficient MEFs during adipogenic induction. A and B , representative immunofluorescent images of Smo or BBS1 that were localized at the primary cilia (acetylated tubulin; green ), in WT, Rab8a −/− , Rab8b KD , Rab8a −/− ;Rab8b KD , or Kif3a −/− MEFs. C and D , representative immunofluorescent images of Fzd2 ( red ) in MEFs before or 24 h after cilia induction. A diagram is used to summarize the findings in WT cells: Fzd2 appeared as a cilium-associated patch before induction; after induction, numerous Fzd2 puncta or vesicles appeared ( red dots ). White arrowheads point to increased Fzd2 puncta that were not associated with a cilium in Rab8a −/− and Rab8a −/− ;Rab8b KD MEFs before induction. Open white arrowheads point to cilia that were not associated with a patch of Fzd2. E , quantification of the percentage of cells with Fzd2 localized to the base of the primary cilium at 0 ( blue bar ) or 24 h ( red bar ) after cilia induction. F , western blots for Fzd2, Tcf1, and β-catenin were performed on total lysates of WT or Rab8-deficient MEFs under uninduced or induced conditions. Full-length Fzd2 was marked by a solid black arrowhead ; truncated Fzd2 marked by an empty arrowhead ; cleaved fragment marked by an asterisk . G , fold changes in Tcf1 protein abundance between uninduced and induced MEFs were quantified from three independent experiments. p values were determined by t test. MEF, mouse embryonic fibroblast.

Article Snippet: Rab8b-specific lentiviral shRNA construct targeting against the 3'UTR of mouse Rab8b was constructed by inserting the annealed complementary oligonucleotides (5'-CCGGGCCAAGAACTAACAGAACTTTCCATGGAAAGTTCTGTTAGTTCTTGGCTTTTTG-3' and 5'-AATTCAAAAAGCCAAGAACTAACAGAACTTTCCATGGAAAGTTCTGTTAGTTCTTGGC-3') into pLK0.1 lentiviral vector (Addgene) between AgeI and EcoRI sites.

Techniques: Translocation Assay, Western Blot