rab7a  (OriGene)

Journal: medRxiv

Article Title: Biallelic WDR91 variants cause a neurodevelopmental disorder through impaired endosomal maturation and autophagy dysregulation

doi: 10.64898/2026.04.03.26349989

Figure Lengend Snippet: A. Time-lapse recording of dynamic changes of the early endosomal marker 2×FYVE-mCherry and the late endosomal protein GFP-Rab7 on endosomes in transduced HEK193 cells (left) and duration of the overlap of 2×FYVE and Rab7 on endosomes (right; one-way ANOVA with Kruskal-Wallis multiple comparison test, ** p<0.01, **** p<10 - ; at least 10 endosomes per cell line were analyzed). B. Colocalization of endogenous EEA1 with Rab7 in WDR91 WT and WDR91 Y15N cells: representative confocal images (left) and assessment of Pearson’s coefficient (right; Mann-Whitney test, *** p<10 - ; at least 30 independent cells per cell line were analyzed). C. EEA1 and Rab7 protein expression in transduced HEK293 cells, representative blot of three independent experiments. Individual fluorescence channels were analyzed separately due to distinct background signal. D. Colocalization of WDR91 with endogenous EEA1 or Rab7 in WDR91 WT and WDR91 Y15N cells: representative confocal images (left) and assessment of Pearson’s coefficient (right; 2-way ANOVA with Sidak multiple comparison test, *** p<10 -3 , **** p<10 -4 ; at least 30 independent cells per cell line were analyzed).

Article Snippet: Cells were then saturated using TBS-Tween 0.01% supplemented with 3% goat serum, and stained with a primary anti-human WDR91 Ab (Abcam) and anti-human EEA1 or anti-human Rab7 Abs (Cell Signaling Technologies) and a secondary goat anti-mouse/rabbit IgG mAb coupled to AF488 or AF687 (Thermo Fisher Scientific).

Techniques: Marker, Comparison, MANN-WHITNEY, Expressing, Fluorescence

Journal: Nature communications

Article Title: Rab7a is an enhancer of TPC2 activity regulating melanoma progression through modulation of the GSK3β/β-Catenin/MITF-axis.

doi: 10.1038/s41467-024-54324-9

Figure Lengend Snippet: Fig. 1 | Expression of Rab7a in melanoma versus non-melanoma cancer cells, correlation with TPC2 expression, and interaction of Rab7a with TPC2. a Gene expression profile (qPCR) of Rab7a in human melanoma lines compared to differ- ent non-melanoma cancer lines. Error bars represent mean values ± SEM. Data points represent biological replicates. b Protein levels corroborating mRNA expression data in melanoma lines (compared to the breast cancer line MDA-MB- 231). Shown are biological replicates, each. c Gene expression profile (qPCR) of Rab7b in human melanoma lines compared to different non-melanoma cancer lines. Data points show biological replicates. Error bars represent mean values ± SEM. Data points represent biological replicates. d Melanoma lines showing strong expression correlation between Rab7a and TPC2 but not TPC1. e Gene expression profile (qPCR) showing relative expression of TPCs in human melanoma lines

Article Snippet: The following antibodies were used for human Rab7 protein from Cell Signaling Technology: 2094S and 9367S, binding around residues Asp193 and Glu188, respectively, corresponding to Exon 6.

Techniques: Expressing, Gene Expression

Journal: Nature communications

Article Title: Rab7a is an enhancer of TPC2 activity regulating melanoma progression through modulation of the GSK3β/β-Catenin/MITF-axis.

doi: 10.1038/s41467-024-54324-9

Figure Lengend Snippet: Fig. 3 | Effect of Rab7 inhibitor on TPC2 activity and physical interaction of Rab7a with TPC2. a, b Inhibition of PI(3,5)P2 evoked currents in endolysosomes (EL), expressing hTPC2 alone or with Rab7a, using the Rab7-inhibitor CID1067700. Shown are representative current density-voltage relationships of enlarged EL, expressing hTPC2WT + hRab7WT or hTPC2WT alone, activated with 1 µM PI(3,5)P2 followed by application of CID1067700 (diff. conc.) and 1 mM ATP (max. effect). c Statistical summary of data as shown in (a, b) at −100 mV. Each dot represents a single current density value measured from one EL. Data were tested for statistical significance with one-way ANOVA test followed by Tukey’s post-test (***p < 0.001, ****p < 0.0001, n = 3). d Cartoon showing CRISPR/Cas9 strategy to knockout TPCN2 in the SK-MEL-5 cell line. e qPCR data showing relative expression of TPC2 in WT and TPC2 KO SK-MEL-5 (n = 3). f Statistical summary of data (average current densities at −100 mV) as shown in (g) (n = 6). g Representative current density-voltage relationships from −100 to +100 mV showing basal, TPC2-A1-P (20 µM) activated and ATP (1 mM) blocked currents.

Article Snippet: The following antibodies were used for human Rab7 protein from Cell Signaling Technology: 2094S and 9367S, binding around residues Asp193 and Glu188, respectively, corresponding to Exon 6.

Techniques: Activity Assay, Inhibition, Expressing, CRISPR, Knock-Out

Journal: Nature communications

Article Title: Rab7a is an enhancer of TPC2 activity regulating melanoma progression through modulation of the GSK3β/β-Catenin/MITF-axis.

doi: 10.1038/s41467-024-54324-9

Figure Lengend Snippet: Fig. 6 | Expression of MITF and GSK3β in different melanoma lines and effects of Rab7a or TPC2 KOor small molecule blockers. a Representative Western blots for MITF and Rab7 protein expression in different melanoma lines and in the breast cancer line MDA-MB-231, normalized to Vinculin. b, c Correlation plot for MITF/Rab7 expression (b) and statistical analysis of experiments as shown in (a) (mean values ± SEM, n = 5–8). Data points represent biological replicates (c). d Representative images of sections from healthy lymphnode (male, abdomen) and melanoma lymphnode metastasis (male, iliacal) samples stained with hMITF antibody (IHC). Scale bars = 5 mm. e IHC evaluation was carried out considering the percentage of stained tumor cells. Statistical significance was assessed by two- tailed unpaired t-test, *p = 0.0125 (mean ± SD). One dot corresponds to one independent human donor (n = 10 for each condition). Genetic knockout of either Rab7a (f, g) or TPC2 (h, i) in SK-MEL-5 cells shows reduction in the protein levels of

Article Snippet: The following antibodies were used for human Rab7 protein from Cell Signaling Technology: 2094S and 9367S, binding around residues Asp193 and Glu188, respectively, corresponding to Exon 6.

Techniques: Expressing, Western Blot, Staining, Two Tailed Test, Knock-Out