rab5a  (Proteintech)

Journal: The Journal of Biological Chemistry

Article Title: Rab5 nucleotide binding promotes oxidative metabolism to fuel hepatocellular carcinoma cell proliferation

doi: 10.1016/j.jbc.2026.111321

Figure Lengend Snippet: Rab5 is upregulated in HCC and associated with cancer hallmarks . A , volcano plot from TNMplot showing expression of known lipid droplet (LD)–associated proteins, including Rab5A/B/C, in HCC tumors versus adjacent normal tissues from n = 53 patients. B , representative immunohistochemistry images from a liver tissue microarray showing Rab5 staining in normal liver (n = 4) versus HCC specimens (n = 7). C , quantification of Rab5 immunohistochemistry scores from the tissue microarray (mean ± SD, ∗∗∗ p < 0.0002; Welch's t test). D , cancer hallmark gene set enrichment analysis based on the study by Menyhart et al . , showing Rab5-associated genes enriched in proliferation, invasion, and evading growth suppressors. E , Kaplan–Meier survival analysis from the TCGA–LIHC dataset stratified by Rab5 expression. High Rab5 expression correlates with poorer overall survival (log-rank test, p = 0.06). HCC, hepatocellular carcinoma; LIHC, Liver Hepatocellular Carcinoma Cohort; TCGA, The Cancer Genome Atlas.

Article Snippet: Antibodies against Rab5A (2143S), PLIN2 (95109), and EEA1 (2411S) were purchased from Cell Signaling Technology.

Techniques: Expressing, Immunohistochemistry, Microarray, Staining

Journal: The Journal of Biological Chemistry

Article Title: Rab5 nucleotide binding promotes oxidative metabolism to fuel hepatocellular carcinoma cell proliferation

doi: 10.1016/j.jbc.2026.111321

Figure Lengend Snippet: Enhanced recruitment of Rab5 to lipid droplets (LDs) upon lipophagy stimulation . A , Western blot analysis of Rab5 and EEA1 GTP-pulldown in regular medium (CT) or 4 h HBSS starvation in Hep3B cells. B , quantification of the fold change of GTP-bound/total Rab5 as well as EEA1 from n = 4 independent experiments. C , quantification of total Rab5–β-actin confirming that starvation does not change overall Rab5 expression. D , Western blot analysis of LDs isolated from Hep3B cells by density gradient centrifugation showing abundant levels of Rab5 in isolated LDs following 4 h of HBSS starvation compared with the regular medium (CT). E , quantification of Rab5–Plin2 in the biochemically isolated LDs. F , fluorescence micrograph showing immunofluorescence staining of Hep3B cells expressing Myc-tagged Rab5 ( green ). Cells were treated with oleic acid for 16 h and then either maintained in regular medium (control) or subjected to 4 h of HBSS starvation before being fixed and stained with Oil Red O (ORO; red ) to label LDs. Note the marked increase in Rab5 staining around LDs in HBSS-starved cells versus control ( yellow arrowheads ). Graphs represent mean ± SD from n = 3 to 4 independent experiments. Statistical significance was determined using an unpaired two-tailed t test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. CT, control; EEA1, early endosome antigen 1; HBSS, Hank’s balanced salt solution; Plin2, perilipin-2.

Article Snippet: Antibodies against Rab5A (2143S), PLIN2 (95109), and EEA1 (2411S) were purchased from Cell Signaling Technology.

Techniques: Western Blot, Expressing, Isolation, Gradient Centrifugation, Fluorescence, Immunofluorescence, Staining, Control, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: Rab5 nucleotide binding promotes oxidative metabolism to fuel hepatocellular carcinoma cell proliferation

doi: 10.1016/j.jbc.2026.111321

Figure Lengend Snippet: Inhibition of Rab5 GTPase activity reduces lipid droplet (LD) catabolism via its GTPase activity . A , Western blot analysis of Rab5 and EEA1 GTP-pulldown in Hep3B cells treated with DMSO (CT) or NAP (100 μM, 48 h). B , quantification of GTP/total Rab5 as well as EEA1 from n = 3 independent experiments. C , confocal micrograph showing ORO-stained LDs ( red ) and DAPI-stained nuclei from Hep3B cells treated with DMSO (control) and NAP (100 μM). D , bar graphs depict quantification of LD number/cell, total LD area/cell, and LD size. E , confocal micrograph showing ORO-stained LDs (red ) and DAPI-stained nucleus from Hep3B cells treated with DMSO (control) and NAP (100 μM) with oleic acid (OA, 150 μM). F , bar graphs depict quantification of LD number/cell, total LD area/cell, and LD size. G , cartoon depicting NAP alteration of Rab5 GTP binding. Graphs represent mean ± SD from n = 3 independent experiments. Statistical significance was determined using an unpaired two-tailed t test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. CT, control; DAPI, 4′,6-diamidino-2-phenylindole; DMSO, dimethyl sulfoxide; EEA1, early endosome antigen 1; NAP, neoandrographolide; ORO, Oil Red O.

Article Snippet: Antibodies against Rab5A (2143S), PLIN2 (95109), and EEA1 (2411S) were purchased from Cell Signaling Technology.

Techniques: Inhibition, Activity Assay, Western Blot, Staining, Control, Binding Assay, Two Tailed Test