uacc62 cells (AcceGen Biotechnology)
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AcceGen Biotechnology
uacc62 cells
Uacc62 Cells, supplied by AcceGen Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/uacc62 cells/product/AcceGen Biotechnology
Average 92 stars, based on 1 article reviews
Uacc62 Cells, supplied by AcceGen Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/uacc62 cells/product/AcceGen Biotechnology
Average 92 stars, based on 1 article reviews
uacc62 cells - by Bioz Stars,
2026-03
92/100 stars
Images
1) Product Images from "RhoA activation promotes glucose uptake to elevate proliferation in MAPK inhibitor resistant melanoma cells"
Article Title: RhoA activation promotes glucose uptake to elevate proliferation in MAPK inhibitor resistant melanoma cells
Journal: bioRxiv
doi: 10.1101/2024.01.09.574940
Figure Legend Snippet: (A) Flow chart to generate MAPKi-resistant melanoma cells. The naïve B-Raf V600E mutant melanoma cells upon exposure to MAPKi show an initial reduction in proliferation. But upon exposure to MAPKi for ∼2 months results in acquisition of resistance. These resistant cells were then cultured in the presence of MAPKi. (B) A375, M481 and UACC62 naïve and resistant cells were compared for proliferation using an imaging based EdU incorporation assay in 2D cell culture conditions. The naïve cells were treated with vehicle control or with MAPKi and the proliferation compared to MAPKi-resistant melanoma cells. The % Proliferation was determined as the ratio of EdU-positive cells to the total cells determined by DAPI counterstain. The scale bar on the image is 50µm. (C) A375 naïve and resistant cells were cultured on glass coverslips. The naive cells were either treated with vehicle control or MAPKi. All the cells were then fixed and stained with phalloidin to evaluate F-actin structures. The scale bar on the image is 10µm. For (B) Three biological repeats were performed with a minimum of 9 images acquired per sample per repeat. The statistical analysis was performed using one-way ANOVA. For (C) Imaging was performed for three biological repeats with a minimum of 10 images acquired per sample per biological repeat. For the statistical significance; ns P ≥ 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. The data represented in (B) is mean ± s.e.m.
Techniques Used: Mutagenesis, Cell Culture, Imaging, Control, Staining
Figure Legend Snippet: (A) A375, M481, UACC62, WM164 and 451Lu were evaluated for the metabolic state. The resistant melanoma cells were compared to naïve cells with vehicle control or MAPKi treatment and seahorse assay performed to evaluate ECAR (readout of glycolysis) and OCR (readout for oxidative phosphorylation). The metabolic rates, ECAR and OCR were normalized to per cell through fixation, DAPI staining and cell count using ImageJ after seahorse assay. (B) Glucose uptake in naïve and resistant A375 melanoma cells was evaluated using NBD-glucose. The cells were then imaged, and the image intensity was quantified per cell using ImageJ. (C) To evaluate the role of F-actin structures in metabolism the A375 resistant melanoma cells were treated with Latrunculin-A to evaluate actin depolymerization. (D) Seahorse assay was then performed for the resistant cells treated with Latrunculin-A after 4h of treatment. (E) A375 naïve cells were overexpressed with a PFKP-GFP followed by seahorse assay. All experiments were performed with 3 biological repeats. For (A) A375, M481, and WM164 had 9 data points per repeat. The UACC62 and 451Lu had atleast 3 datapoint per repeat. For (B) Each biological sample in the repeat had at least 12 images. (D) and (E) had 3 data points per repeat. The statistical analysis was performed using one-way ANOVA for (A), (B) and (D). Welch’s t-test was used for (E). For the statistical significance; ns P ≥ 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. The data represented in (B) is mean ± s.e.m.
Techniques Used: Control, Phospho-proteomics, Staining, Cell Counting
Figure Legend Snippet: (A) RhoA activation was evaluated in A375, UACC62 and M481 naïve and resistant melanoma cells using the RhoA activation pull down assay. The band intensities were quantified for the rho pull down as a ratio of resistant to naïve cells. (B) The A375 naïve and resistant cells were treated with Y27632 and GSK269962A for 48h followed by EdU incorporation assay to evaluate proliferation. (C) A375 naïve and resistant melanoma cells treated with GSK269962A were evaluated for glucose uptake using NBD-glucose and the intensities quantified per cells using ImageJ. (D) Metabolic rate was quantified for A375 resistant melanoma cells treated with GSK269962A and evaluated for ECAR and OCR. (E) RhoA was activated in naïve A375 and M481 using a constitutively active RhoA (CA-RhoA) and evaluated for glucose uptake using NBD-glucose assay and image intensities quantified per cell which was compared to empty vector (EV). (F) Seahorse assay was performed to evaluate ECAR and OCR in naïve A375 and M481 melanoma cells with EV or CA-RhoA. (G) Proliferation between EV and CA-RhoA was evaluated using EdU incorporation assay for the naïve A375 and M481 melanoma cells. For (A) 4 biological repeats, (B) Three biological repeats were performed with three data points per repeat, (C) (E) and (G) Three biological repeats with at least 10 images per sample per repeat, (D) Three biological repeats with four data points per sample per repeat. (F) Three biological repeats with six data points per repeat. The statistical analysis was performed using one-way ANOVA for (C), and (D). Welch’s t-test was used for (E), (F) and (G). For the statistical significance; ns P ≥ 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. The data represented in (B) is mean ± s.e.m.
Techniques Used: Activation Assay, Pull Down Assay, Glucose Assay, Plasmid Preparation
Figure Legend Snippet: (A) The activity of PKA in A375, M481 and UACC62 naïve and resistant melanoma cells was evaluated using the PKA activation assay. (B) To induce PKA activation in A375 resistant cells 8-Bromo-cAMP was used and the metabolic state was evaluated using seahorse assay. (C) Proliferation was evaluated in A375 resistant melanoma cells upon 8-Bromo-cAMP treatment using EdU incorporation assay. For (A) each data point represents a biological repeat. For (B) Three biological repeats were performed with four data points per repeat. For (C) three biological repeats ten data points per repeat. The statistical analysis was performed using one-way ANOVA for (B) and (C). Welch’s t-test was used for (A). For the statistical significance; ns P ≥ 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. The data represented in (B) is mean ± s.e.m.
Techniques Used: Activity Assay, Activation Assay
