Journal: BMC Cancer

Article Title: Regulation of EMT-MET and chemoresistance by the Lc3Cer-synthase B3GNT5

doi: 10.1186/s12885-025-14717-5

Figure Lengend Snippet: Increased expression and copy number amplifications of B3GNT5 are highly prevalent in human cancers. a The TNMplot online database was queried for the expression of B3GNT5 in the indicated cancers. Cancers where tumor levels of B3GNT5 were significantly higher than in normal tissues are marked with an asterisk (*). b For the indicated cancers, TCGA data was analyzed for B3GNT5 (ENSG00000176597) genomic alterations. Plots b-g show the percentage of cases altered and colored by CNV gain alterations (red segments), CNV loss alterations (green segments), and simple somatic mutations (SSM, blue segments). c – g TCGA data for the indicated cancers was analyzed for CNV gain alterations in genes of sphingolipid, lactoside, neolactoside, ganglioside, isogloboside, and globosides metabolic pathways

Article Snippet: Antibodies used for immunoblotting were diluted in a 2% solution of bovine serum albumin (MilliporeSigma, 126609) dissolved in 50 mM Tris-Cl, pH 7.5, 150 mM NaCl, 0.1% Tween-20 and were: B3GNT5 (1:1000, 20422-1-AP, Proteintech), MRP1/ABCC1 (1:2000, Cell Signaling Technology, 72202), MDR1/ABCB1 (1:2000, Cell Signaling Technology 13342) GAPDH (1:3000, 2118, Cell Signaling Technology, Danvers, MA), vimentin (1:1000, Cell Signaling Technology, 5741), N-cadherin (1:1000, Cell Signaling Technology, 13116), β-Catenin (1:1000, Cell Signaling Technology, 8480), SNAIL (1:1000, Cell Signaling Technology, 3879), SLUG (1:1000, Cell Signaling Technology, 9585), ZEB1 (1:1000, Cell Signaling Technology, 3396), E-Cadherin (1:1000, Cell Signaling Technology, 3195), pan-keratin (1:1000, Cell Signaling Technology, 4545).

Techniques: Expressing

Journal: BMC Cancer

Article Title: Regulation of EMT-MET and chemoresistance by the Lc3Cer-synthase B3GNT5

doi: 10.1186/s12885-025-14717-5

Figure Lengend Snippet: Increased expression of B3GNT5 is significantly associated with poor outcomes in human cancers. a – i The Human Protein Atlas online tool was used to generate Kaplan-Meier plots and analyze the outcomes of patients with high and low tumor expression of B3GNT5 . j , k Kaplan-Meier plots generated using pan-cancer TCGA genomic data for cases in which the B3GNT5 (ENSG00000176597) loci was altered (any type of mutation) or unaltered (no mutation in B3GNT5 ) and the indicated outcome measures for overall survival (OS; panel j ) or disease-free survival (DFS; panel k) determined (Prism 8)

Article Snippet: Antibodies used for immunoblotting were diluted in a 2% solution of bovine serum albumin (MilliporeSigma, 126609) dissolved in 50 mM Tris-Cl, pH 7.5, 150 mM NaCl, 0.1% Tween-20 and were: B3GNT5 (1:1000, 20422-1-AP, Proteintech), MRP1/ABCC1 (1:2000, Cell Signaling Technology, 72202), MDR1/ABCB1 (1:2000, Cell Signaling Technology 13342) GAPDH (1:3000, 2118, Cell Signaling Technology, Danvers, MA), vimentin (1:1000, Cell Signaling Technology, 5741), N-cadherin (1:1000, Cell Signaling Technology, 13116), β-Catenin (1:1000, Cell Signaling Technology, 8480), SNAIL (1:1000, Cell Signaling Technology, 3879), SLUG (1:1000, Cell Signaling Technology, 9585), ZEB1 (1:1000, Cell Signaling Technology, 3396), E-Cadherin (1:1000, Cell Signaling Technology, 3195), pan-keratin (1:1000, Cell Signaling Technology, 4545).

Techniques: Expressing, Generated, Mutagenesis

Journal: BMC Cancer

Article Title: Regulation of EMT-MET and chemoresistance by the Lc3Cer-synthase B3GNT5

doi: 10.1186/s12885-025-14717-5

Figure Lengend Snippet: Partial depletion of B3GNT5 resulted in the accumulation of GlcCer and LacCer. a , b B3GNT5 was targeted in HeLa cells using CRISPR-Cas9. An isolated clone was analyzed using immunoblotting with the indicated antibodies. Blots were quantified by densitometry using ImageJ 1.53. The y-axis shows band intensity normalized to GAPDH and expressed as fold change relative to wild-type. n = 5. c , d Mass spectrometry analysis of the indicated lipids per mg of input protein in HeLa/HeLa B3GNT5 PKO cells. n = 3. Bar graphs with mean ± SEM shown; T-test analysis (Prism 8): ns, not significant; *, P ≤ 0.05; **, P ≤ 0.01; ****, P ≤ 0.0001

Article Snippet: Antibodies used for immunoblotting were diluted in a 2% solution of bovine serum albumin (MilliporeSigma, 126609) dissolved in 50 mM Tris-Cl, pH 7.5, 150 mM NaCl, 0.1% Tween-20 and were: B3GNT5 (1:1000, 20422-1-AP, Proteintech), MRP1/ABCC1 (1:2000, Cell Signaling Technology, 72202), MDR1/ABCB1 (1:2000, Cell Signaling Technology 13342) GAPDH (1:3000, 2118, Cell Signaling Technology, Danvers, MA), vimentin (1:1000, Cell Signaling Technology, 5741), N-cadherin (1:1000, Cell Signaling Technology, 13116), β-Catenin (1:1000, Cell Signaling Technology, 8480), SNAIL (1:1000, Cell Signaling Technology, 3879), SLUG (1:1000, Cell Signaling Technology, 9585), ZEB1 (1:1000, Cell Signaling Technology, 3396), E-Cadherin (1:1000, Cell Signaling Technology, 3195), pan-keratin (1:1000, Cell Signaling Technology, 4545).

Techniques: CRISPR, Isolation, Western Blot, Mass Spectrometry

Journal: BMC Cancer

Article Title: Regulation of EMT-MET and chemoresistance by the Lc3Cer-synthase B3GNT5

doi: 10.1186/s12885-025-14717-5

Figure Lengend Snippet: Reduction of B3GNT5 levels significantly increased the chemoresistance of HeLa cells to vinorelbine and paclitaxel. a – d HeLa and HeLa B3GNT5 PKO cells were treated with the indicated drugs and concentrations for 72 h, and viability assayed with WST-8. n = 3. e , f HeLa and HeLa B3GNT5 PKO cells were seeded at low density in 10 cm tissue culture plates, allowed to attach for 18 h, and then treated with the indicated chemotherapeutic agents for 72 h. The media was then exchanged, and cells allowed to grow until colonies contained > 200 cells. Cells were then fixed, the colonies stained, and colonies with > 200 cells counted. n = 3. g) Wild-type and B3GNT5 PKO cells were analyzed by immunoblotting for the indicated antibodies, and quantified by densitometry using ImageJ 1.53. The y-axis shows band intensity normalized to GAPDH and expressed as fold change relative to wild type. n = 5. All cells were grown at 37 °C and 5% CO 2 . X-axis in panels a-d is in log2 scale. T-test analysis (panels e, f), IC50 fitting, and ANOVA (panels a-d) were performed with Prism 8; ns, not significant; *, P ≤ 0.05; **, ≤ 0.01, P ***, P ≤ 0.005; ****, P ≤ 0.0001

Article Snippet: Antibodies used for immunoblotting were diluted in a 2% solution of bovine serum albumin (MilliporeSigma, 126609) dissolved in 50 mM Tris-Cl, pH 7.5, 150 mM NaCl, 0.1% Tween-20 and were: B3GNT5 (1:1000, 20422-1-AP, Proteintech), MRP1/ABCC1 (1:2000, Cell Signaling Technology, 72202), MDR1/ABCB1 (1:2000, Cell Signaling Technology 13342) GAPDH (1:3000, 2118, Cell Signaling Technology, Danvers, MA), vimentin (1:1000, Cell Signaling Technology, 5741), N-cadherin (1:1000, Cell Signaling Technology, 13116), β-Catenin (1:1000, Cell Signaling Technology, 8480), SNAIL (1:1000, Cell Signaling Technology, 3879), SLUG (1:1000, Cell Signaling Technology, 9585), ZEB1 (1:1000, Cell Signaling Technology, 3396), E-Cadherin (1:1000, Cell Signaling Technology, 3195), pan-keratin (1:1000, Cell Signaling Technology, 4545).

Techniques: Staining, Western Blot

Journal: BMC Cancer

Article Title: Regulation of EMT-MET and chemoresistance by the Lc3Cer-synthase B3GNT5

doi: 10.1186/s12885-025-14717-5

Figure Lengend Snippet: HeLa B3GNT5 PKO cells had significantly reduced invasive capacity. a Representative images of the invasive capacity of HeLa and HeLa B3GNT5 PKO cells as examined by quantifying the number of cells that migrated through a Matrigel matrix and were stained with crystal violet. Before assays, cells were serum-starved for 2 h prior to seeding in the upper chamber of a transwell insert containing Matrigel. Cells were allowed to grow for 48 h at 37 °C and 5% CO 2 . The lower chamber contained media containing 10% FBS. Plot is bar graph with mean and ± SEM. T-test analysis (Prism 8): ***, P ≤ 0.005; n = 3

Article Snippet: Antibodies used for immunoblotting were diluted in a 2% solution of bovine serum albumin (MilliporeSigma, 126609) dissolved in 50 mM Tris-Cl, pH 7.5, 150 mM NaCl, 0.1% Tween-20 and were: B3GNT5 (1:1000, 20422-1-AP, Proteintech), MRP1/ABCC1 (1:2000, Cell Signaling Technology, 72202), MDR1/ABCB1 (1:2000, Cell Signaling Technology 13342) GAPDH (1:3000, 2118, Cell Signaling Technology, Danvers, MA), vimentin (1:1000, Cell Signaling Technology, 5741), N-cadherin (1:1000, Cell Signaling Technology, 13116), β-Catenin (1:1000, Cell Signaling Technology, 8480), SNAIL (1:1000, Cell Signaling Technology, 3879), SLUG (1:1000, Cell Signaling Technology, 9585), ZEB1 (1:1000, Cell Signaling Technology, 3396), E-Cadherin (1:1000, Cell Signaling Technology, 3195), pan-keratin (1:1000, Cell Signaling Technology, 4545).

Techniques: Staining

Journal: BMC Cancer

Article Title: Regulation of EMT-MET and chemoresistance by the Lc3Cer-synthase B3GNT5

doi: 10.1186/s12885-025-14717-5

Figure Lengend Snippet: Decreased expression of B3GNT5 altered the expression of EMT-MET markers. The indicated proteins in HeLa and HeLa B3GNT5 PKO cells were analyzed by western blotting, and representative immunoblots ( a , c ) and corresponding densitometry ( b , d ) analysis shown. n = 3. Plots in panels b and d are bar graphs with mean and ± SEM, with y-axis showing the relative band intensity normalized to the wild-type. Blots were quantified with ImageJ 1.53. T-test (Prism 8): ns, not significant; *, P ≤ 0.05; ****, P ≤ 0.0001

Article Snippet: Antibodies used for immunoblotting were diluted in a 2% solution of bovine serum albumin (MilliporeSigma, 126609) dissolved in 50 mM Tris-Cl, pH 7.5, 150 mM NaCl, 0.1% Tween-20 and were: B3GNT5 (1:1000, 20422-1-AP, Proteintech), MRP1/ABCC1 (1:2000, Cell Signaling Technology, 72202), MDR1/ABCB1 (1:2000, Cell Signaling Technology 13342) GAPDH (1:3000, 2118, Cell Signaling Technology, Danvers, MA), vimentin (1:1000, Cell Signaling Technology, 5741), N-cadherin (1:1000, Cell Signaling Technology, 13116), β-Catenin (1:1000, Cell Signaling Technology, 8480), SNAIL (1:1000, Cell Signaling Technology, 3879), SLUG (1:1000, Cell Signaling Technology, 9585), ZEB1 (1:1000, Cell Signaling Technology, 3396), E-Cadherin (1:1000, Cell Signaling Technology, 3195), pan-keratin (1:1000, Cell Signaling Technology, 4545).

Techniques: Expressing, Western Blot

Journal: BMC Cancer

Article Title: Regulation of EMT-MET and chemoresistance by the Lc3Cer-synthase B3GNT5

doi: 10.1186/s12885-025-14717-5

Figure Lengend Snippet: HeLa with decreased B3GNT5 expression had significantly reduced activation of RTKs upon stimulation with FBS. a HeLa and HeLa B3GNT5 PKO cells were cultured in media lacking fetal bovine serum (FBS) for 3 h, and then incubated with media containing 10% FBS for 30 min. Extracts were run in triplicate. Shown are representative image of Proteome Profiler Human RTK arrays. Spots outlined are shown in panel b. Shown are also immunoblots and quantification with the indicated antibody of input extracts used to incubate RTK arrays. n = 3. b Spots from panel a that were quantification using densitometry (ImageJ 1.53), with data shown as relative normalized to the wild-type and plotted in panel c. T-tests were performed with Prism 8. ns, not significant; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.0005

Article Snippet: Antibodies used for immunoblotting were diluted in a 2% solution of bovine serum albumin (MilliporeSigma, 126609) dissolved in 50 mM Tris-Cl, pH 7.5, 150 mM NaCl, 0.1% Tween-20 and were: B3GNT5 (1:1000, 20422-1-AP, Proteintech), MRP1/ABCC1 (1:2000, Cell Signaling Technology, 72202), MDR1/ABCB1 (1:2000, Cell Signaling Technology 13342) GAPDH (1:3000, 2118, Cell Signaling Technology, Danvers, MA), vimentin (1:1000, Cell Signaling Technology, 5741), N-cadherin (1:1000, Cell Signaling Technology, 13116), β-Catenin (1:1000, Cell Signaling Technology, 8480), SNAIL (1:1000, Cell Signaling Technology, 3879), SLUG (1:1000, Cell Signaling Technology, 9585), ZEB1 (1:1000, Cell Signaling Technology, 3396), E-Cadherin (1:1000, Cell Signaling Technology, 3195), pan-keratin (1:1000, Cell Signaling Technology, 4545).

Techniques: Expressing, Activation Assay, Cell Culture, Incubation, Western Blot

Journal:

Article Title: Apoptosis and Autophagy After Mitochondrial or Endoplasmic Reticulum Photodamage †

doi: 10.1111/j.1751-1097.2007.00088.x

Figure Lengend Snippet: Western blots showing processing of the LC3-I protein to the faster-migrating LC3-II species in L1210 vs L1210/Atg7− cells. Analyses are of control cultures (top), and cultures harvested 30 min or 24 h after administration of low- or high-dose PDT with MC (left) or CPO (right).

Article Snippet: An antibody to the murine LC3 protein was provided by Proteintech Group, Inc. (Chicago, IL).

Techniques: Western Blot, Control