Journal: Cells

Article Title: 2,3-Bisphosphoglycerate Mutase (BPGM), a Metabolic Player Shaping Stress-Adaptive Transcriptional States in Clear Cell Renal Cell Carcinoma

doi: 10.3390/cells15070633

Figure Lengend Snippet: BPGM expression is elevated in human clear cell renal cell carcinoma. ( a ) Representative immunoblot showing BPGM protein levels in ccRCC tumor tissue of four representative patients (P1–4) and matched adjacent normal kidney samples. ( b ) Paired analysis of BPGM protein expression in matched control and tumor samples from individual patients ( n = 61 paired samples). Each data point represents one biological replicate (individual patient sample). ( c ) Violin plot depicting the distribution of BPGM expression levels in normal kidney tissue and ccRCC samples, illustrating increased expression and interindividual variability in tumor tissue. The dashed line indicates median and dotted lines the interquartile range. Statistical analysis was performed using a paired Student’s t -test. Exact p -values are indicated in the figure.

Article Snippet: BPGM protein was detected using a polyclonal rabbit anti-BPGM antibody (catalog #NBP1-86064, Novus Biologicals, Centennial, CO, USA).

Techniques: Expressing, Western Blot, Control

Journal: Cells

Article Title: 2,3-Bisphosphoglycerate Mutase (BPGM), a Metabolic Player Shaping Stress-Adaptive Transcriptional States in Clear Cell Renal Cell Carcinoma

doi: 10.3390/cells15070633

Figure Lengend Snippet: ccRCC cells show limited sensitivity to epigenetic stress and elevated BPGM expression. ( a , b ) Crystal violet assays of ( a ) 2102EP and ( b ) A498 cells after 24 h treatment with increasing concentrations of Vorinostat (VS; 2–8 µM). ( a ) n = 10 per condition. ( b ) n = 10 (control, 2 µM, 8 µM) and n = 9 (3 µM, 4 µM; one outlier removed). ( c , d ) Basal BPGM protein expression in 2102EP and A498 cells shown as representative immunoblot ( c ) and quantification ( d ) ( n = 6 per condition). ( e ) BPGM mRNA expression after 24 h VS treatment in 2102EP and A498 cells. Expression is shown relative to control. n = 6 (2102EP); n = 5 (A498 control; one outlier removed) and n = 6 (A498 VS). Data are presented as box plots (median with interquartile range; whiskers indicate min–max). Each data point represents one biological replicate. Statistical analysis was performed using Brown–Forsythe ANOVA with Dunnett’s T3 multiple comparisons test ( a , b ), Welch’s t -test ( d ), and unpaired Student’s t -test (2102EP) or Welch’s t -test (A498) for ( e ). Details on test selection and normality assessment are provided in Methods. For clarity, only step-wise comparisons along the concentration series are displayed in panels ( a , b ). Complete multiple comparisons are provided in . Color code: white: 2102EP control; beige: A498 control; red: 2102EP + vorinostat; blue: A498 + vorinostat. Exact p -values are indicated in the figure.

Article Snippet: BPGM protein was detected using a polyclonal rabbit anti-BPGM antibody (catalog #NBP1-86064, Novus Biologicals, Centennial, CO, USA).

Techniques: Expressing, Control, Western Blot, Selection, Concentration Assay

Journal: PLoS ONE

Article Title: Novel PDE4 Inhibitors Derived from Chinese Medicine Forsythia

doi: 10.1371/journal.pone.0115937

Figure Lengend Snippet: A. 5×10 5 RAW264.7 cells were seeded in 96 wells for 18 h. Cells were primed with compounds at different concentration for 3 h before treated with LPS (1 ng/ml) for additional 8 h. TNF cytokine releases were monitored by ELISA. B. PBMC (0.2 ml at 1×10 5 /ml) were primed with compounds at different concentration for 3 h before treated with LPS (1 ng/ml) for additional 8 h. TNF cytokine releases were monitored by ELISA. % of TNF secretion were calculated and graphed. C. Summary of compound IC 50 . The data represent n = 3–6 experiments.

Article Snippet: IL1β, TNFα, IL6 mouse ELISA kit, human TNFα were from R&D systems.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Military Medical Research

Article Title: Endophilin A2 controls touch and mechanical allodynia via kinesin-mediated Piezo2 trafficking.

doi: 10.1186/s40779-024-00520-z

Figure Lengend Snippet: Fig. 9 Hypothetical model illustrating that endophilin A2 (EndoA2) interacts with Piezo2 and KIF5B to form a complex to regulate the membrane trafficking of Piezo2 in sensory neurons and thus contributes to mechanical hypersensitivity

Article Snippet: DRG neurons from L4–L6 of mice were cultured for 2 d. Following fixation, DRG coverslips were treated with primary antibodies against Piezo2 (rabbit, 1:200, Novus, USA, NBP1-78,624) and NF200 (mouse, 1:200, Sigma, USA, N0142) and the appropriate secondary antibodies.

Techniques: Membrane

Journal: BMC cancer

Article Title: The Hippo pathway promotes platinum-based chemotherapy by inhibiting MTF1-dependent heavy metal response.

doi: 10.1186/s12885-025-13661-8

Figure Lengend Snippet: Fig. 1 Platinum-based chemotherapy drugs activate MTF1 and the heavy metal response. A Illustration of the chemical structures for platinum-based compounds cisplatin, carboplatin and oxaliplatin. Platinum (Pt) is labeled in red. B Platinum-based compounds induce MTF1’s nuclear translocation. HEK293A cells were treated with CdCl2 (50 µM), cisplatin (50 µM), carboplatin (200 µM) and oxaliplatin (100 µM) for 4 h and subjected to immunofluorescent staining. Scale bar, 20 µm. C-D Platinum-based compounds promote the transcription of heavy metal response genes. The transcription of heavy metal response genes MT1A (C) and MT1F (D) was examined in the indicated cell lines by q-PCR (mean ± s.d., n = 3 biological replicates). The indicated cells were treated with cisplatin (50 µM), carboplatin (200 µM) and oxaliplatin (100 µM) for 12 h. * p < 0.05, ** p < 0.01, *** p < 0.001 (Student’s t-test)

Article Snippet: For immunofluorescent staining, anti-MTF1 antibody (NBP1-86,380, 1:200 dilution) was obtained from Novus Biologicals.

Techniques: Labeling, Translocation Assay, Staining

Journal: BMC cancer

Article Title: The Hippo pathway promotes platinum-based chemotherapy by inhibiting MTF1-dependent heavy metal response.

doi: 10.1186/s12885-025-13661-8

Figure Lengend Snippet: Fig. 2 MTF1 and its associated heavy metal response induce resistance to platinum-based chemotherapy. A Validation of the MTF1 knockout (KO) cells by Western blot. B-D Loss of MTF1 inhibits cell viability under cisplatin treatment. The indicated wild-type and the MTF1 KO cells were treated with cisplatin (50 µM for HEK293A, 100 µM for HEY, 100 µM for HEC1A, and 10 µM for CAL-27) for 24 h and visualized by crystal violet staining (B). Wild-type and MTF1 KO HEY (C) and HEC1A (D) cells were treated with the indicated concentration of cisplatin for 24 h and quantified for relative viability (mean ± s.d., n = 3 biological replicates). E–F Loss of MTF1 inhibits tumor growth under cisplatin treatment. Wild-type and MTF1 KO HEY cells were subjected to xenograft study and treated with cisplatin (5 mg/kg) twice a week. The collected tumors at the endpoint were shown (E), and tumor weight was measured and quantified (mean ± s.d., n = 5 mice per group) (F). ** p < 0.01, *** p < 0.001 (Student’s t-test). ns, no significance. G-H Wild-type and MTF1 KO HEC1A cells were subjected to xenograft study and treated with cisplatin (5 mg/kg) twice a week. The collected tumors at the endpoint were shown (E), and tumor weight was measured and quantified (mean ± s.d., n = 5 mice per group) (F). * p < 0.05, *** p < 0.001 (Student’s t-test). ns, no significance. I-J Loss of MTF1 does not affect mitomycin C and camptothecin-induced cell death. Wild-type and MTF1 KO HEY (I) and HEC1A (J) cells were treated with mitomycin C (20 µM) and camptothecin (50 µM) for 24 h, subjected to crystal violet staining, and quantified for relative viability (mean ± s.d., n = 3 biological replicates). * p < 0.05, ** p < 0.01, *** p < 0.001 (Student’s t-test). ns, no significance. K-M Reconstitution of heavy metal response proteins MT1A and MT2A rescues MTF1 KO cell viability under cisplatin treatment. The indicated MTF1 KO cells were reconstituted with MT1A and MT2A (K), treated with cisplatin (10 µM for HEY, 25 µM for HEC1A) for 48 h, and visualized by crystal violet staining (L). Wild-type HEY and HEC1A and their indicated MTF1 KO cells were treated with the indicated concentration of cisplatin for 48 h and quantified for relative viability (mean, n = 3 biological replicates) (M)

Article Snippet: For immunofluorescent staining, anti-MTF1 antibody (NBP1-86,380, 1:200 dilution) was obtained from Novus Biologicals.

Techniques: Biomarker Discovery, Knock-Out, Western Blot, Staining, Concentration Assay

Journal: BMC cancer

Article Title: The Hippo pathway promotes platinum-based chemotherapy by inhibiting MTF1-dependent heavy metal response.

doi: 10.1186/s12885-025-13661-8

Figure Lengend Snippet: Fig. 3 The Hippo-MTF1 pathway modulates platinum-based chemotherapy. A-B Loss of Hippo pathway components LATS1/2, MOB1A/B and NF2 protects cells under cisplatin treatment. The indicated Hippo pathway component knockout cells were treated with cisplatin (50 µM) for 48 h and visualized by crystal violet staining (A). The indicated Hippo pathway component knockout cells were treated with the indicated concentration of cisplatin for 24 h and quantified for relative viability (mean, n = 3 biological replicates) (B). C-D MTF1 is required for the LATS1/2 deficiency-induced cisplatin resistance. The indicated cells were treated with cisplatin (50 µM) for 48 h and visualized by crystal violet staining (C). The indicated cells were treated with the indicated concentration of cisplatin for 24 h and quantified for relative viability (mean, n = 3 biological replicates) (D). E Validation of the MTF1 KO HEK293A cells reconstituted with MTF1 and its indicated mutants. Western blot was performed using the indicated antibodies. F-G Reconstitution of MTF1 S152A mutant rescues the MTF1 KO HEK293A cell viability under the treatment with cisplatin and carboplatin. The MTF1KO HEK293A cells were reconstituted with MTF1 and its indicated mutants, treated with cisplatin (50 µM) and carboplatin (200 µM) for 48 h, visualized by crystal violet staining (F), and quantified for relative viability (mean ± s.d., n = 3 biological replicates) (G). *** p < 0.001 (Student’s t-test). H Validation of the MTF1 KO HEY cells reconstituted with MTF1 and its indicated mutants. Western blot was performed using the indicated antibodies. I-J Reconstitution of MTF1 S152A mutant rescues the MTF1 KO HEY cell viability and xenograft tumor growth under cisplatin treatment. The MTF1KO HEY cells were reconstituted with MTF1 and its indicated mutants, treated with cisplatin (100 µM) for 48 h, visualized by crystal violet staining (I), and quantified for relative viability (mean ± s.d., n = 3 biological replicates) (J). The MTF1 KO HEY cells were reconstituted with MTF1 and its indicated mutants, subjected to xenograft study, and treated with cisplatin (5 mg/kg) twice a week. The collected tumors at the endpoint were shown (K), and tumor weight was measured and quantified (mean ± s.d., n = 5 mice per group) (L). * p < 0.05, *** p < 0.001 (Student’s t-test). ns, no significance

Article Snippet: For immunofluorescent staining, anti-MTF1 antibody (NBP1-86,380, 1:200 dilution) was obtained from Novus Biologicals.

Techniques: Knock-Out, Staining, Concentration Assay, Biomarker Discovery, Western Blot, Mutagenesis

Journal: BMC cancer

Article Title: The Hippo pathway promotes platinum-based chemotherapy by inhibiting MTF1-dependent heavy metal response.

doi: 10.1186/s12885-025-13661-8

Figure Lengend Snippet: Fig. 5 Clinical relevance of the Hippo-MTF1 pathway in platinum-based chemotherapy. A-C Kaplan–Meier curves of overall survival of lung adenocarcinoma (LUAD) patients is stratified by the expression levels of MT1A (A), MT2A (B) and MTF1 (C). The LUAD RNAseq datasets with clinic data were downloaded from the Cancer Genome Atlas (TCGA) data portal. A total of 491 patients were analyzed for their overall survival. The p value was calculated by using the log-rank (Mantel-Cox) test. D-E The expression of heavy metal response genes MT1A and MT2A is positively correlated with that of Hippo signaling downstream genes CTGF and CYR61 (D), but not with that of YAP1 or WWTR1 (E), in the cisplatin/carboplatin-treated LUAD patient samples. The indicated gene expression from a total of 150 patients who were treated with cisplatin and carboplatin were subjected to Spearman correction analysis. Correlation coefficient R value and p value were calculated by GraphPad prism software

Article Snippet: For immunofluorescent staining, anti-MTF1 antibody (NBP1-86,380, 1:200 dilution) was obtained from Novus Biologicals.

Techniques: Expressing, Gene Expression, Software

Journal: eLife

Article Title: A unified view of low complexity regions (LCRs) across species

doi: 10.7554/eLife.77058

Figure Lengend Snippet: ( A ) Distribution of total and distinct LCRs for all LCR-containing proteins in the human proteome. The number in each square is the number of proteins in the human proteome with that number of total and distinct LCRs and is represented by the colorbar. ( B ) Illustration of different protein groups defined by their LCR combinations, and the number and percentage (%) of proteins that fall into each group. Group definitions are mutually exclusive. ( C ) Dotplot and schematic of RPA43. K-rich LCRs are highlighted in blue, and are labeled K1-K3. Sequences of K1-K3 are shown below the schematic. ( D ) Immunofluorescence of HeLa cells transfected with RPA43 constructs. HeLa cells were seeded on fibronectin-coated coverslips and transfected with the indicated GFP-RPA43 constructs, and collected ~48 hr following transfection. DAPI, GFP, and MPP10 channels are shown. Scale bar is 5 μm. ( E ) Droplet formation assays using GFP-fused RPA43 C-terminus in vitro. Droplet assays were performed with 8.3 μM purified protein. Scale bar is 10 μm. See also .

Article Snippet: Antibody , Anti-MPHOSPH10 (MPP10) (rabbit polyclonal) , Novus Biologicals , NBP1-84341 , (1:100).

Techniques: Labeling, Immunofluorescence, Transfection, Construct, In Vitro, Purification

Journal: eLife

Article Title: A unified view of low complexity regions (LCRs) across species

doi: 10.7554/eLife.77058

Figure Lengend Snippet: ( A ) Distribution of total and distinct LCRs for all LCR-containing proteins in the human proteome from , without binning proteins with 10+total LCRs and/or 10+distinct LCRs. The number in each square is the number of proteins in the human proteome with that number of total and distinct LCRs and is represented by the colorbar. ( B ) Disorder tendency (predicted by IUPred2A) of WT or ΔK1,2,3 RPA43. Coordinates of the three K-rich LCRs of RPA43 are indicated in blue. ( C ) Immunofluorescence of RPA43 constructs in HeLa cells. HeLa cells were seeded on fibronectin-coated coverslips and transfected with the indicated GFP-RPA43 constructs, and collected ~48 hr following transfection. DAPI, GFP, and MPP10 channels are shown. Scale bar is 5 μm. The number of K-rich LCRs present and fibrillar center (FC) localization scoring is shown to the right of each construct (‘+++’ to ‘+’=strong FC localization to uniform nuclear localization, ‘-’=nucleolar exclusion).

Article Snippet: Antibody , Anti-MPHOSPH10 (MPP10) (rabbit polyclonal) , Novus Biologicals , NBP1-84341 , (1:100).

Techniques: Immunofluorescence, Construct, Transfection

Journal: eLife

Article Title: A unified view of low complexity regions (LCRs) across species

doi: 10.7554/eLife.77058

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-MPHOSPH10 (MPP10) (rabbit polyclonal) , Novus Biologicals , NBP1-84341 , (1:100).

Techniques: Recombinant, Expressing, Plasmid Preparation, Software

Journal: Journal of Neuropathology and Experimental Neurology

Article Title: Friedreich Ataxia: Developmental Failure of the Dorsal Root Entry Zone

doi: 10.1093/jnen/nlx087

Figure Lengend Snippet: DREZ in a normal control and a patient with FA shown by α-dystroglycan immunohistochemistry and double-label immunofluorescence of periaxin and GFAP. (A, B) Normal control; FA (C, D) (FA2, Table, GAA 1200/1200). (A) In the normal control, compact α-dystroglycan reaction product surrounds the dome of the CNS portion of the DREZ. (B) Double-label immunofluorescence of periaxin and GFAP shows a relatively sharp transition of myelinating Schwann cells (red) to GFAP-reactive CNS tissue (green) in the DREZ. (C) In FA, the demarcation between α-dystroglycan-positive and negative regions of the DREZ is interrupted by short breaks where CNS tissue penetrates into the DR (arrows). (D) In FA, the normal demarcation between Schwann cells (periaxin) and CNS (GFAP) is indistinct. (A, C) α-dystroglycan; (B, D) double-label immunofluorescence of periaxin (red) and GFAP (green). Scale bars: (A, C) 200 µm; (B, D) 100 µm.

Article Snippet: Sources of commercial antibodies; catalog numbers; host species and clonality; final protein concentrations or, in case of ascites fluid, dilution; and antigen retrieval methods were as follows: anti-PMP-22 (Abcam, Cambridge, MA, ab-31851, rabbit polyclonal, 2 μg protein/mL, incubation in 80% ethanol at 4 °C overnight); anti-P0 (Abcam, ab15508, rabbit polyclonal, 1 μg protein/mL, incubation in 80% ethanol at 4 °C overnight, followed by 0.01 M citric acid-sodium citrate buffer, pH 6, for 20 minutes at 95 °C); anti-PLP (Abcam, ab-105784, rabbit polyclonal, 2 μg protein/mL, incubation in 80% ethanol at 4 °C overnight, followed by 0.01 M citric acid-sodium citrate buffer, pH 6, for 20 minutes at 95 °C); anti-α-dystroglycan (Novus Biologicals, Littleton, CO, NBP1-49634, mouse monoclonal, 0.4 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, for 20 minutes at 95 °C); anti-periaxin (Novus, NBP1-89598, rabbit polyclonal, 2 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, 20 minutes at 95 °C); anti-laminin (Sigma-Aldrich, St. Louis, MO, L8271, mouse monoclonal, ascites fluid, diluted 1:1000, proteinase K [Sigma, P-6556], 0.1 μg/mL in tris buffer, pH 8, containing 0.01 M NaCl for 30 minutes at 37 °C); anti-GFAP (BioLegend, San Diego, CA, 837201, mouse monoclonal, ascites fluid, diluted 1:500, DIVA [a proprietary decloaking solution sold by Biocare Medical, Pacheco, CA] 1× for 30 minutes at 95 °C); anti-vimentin (Santa Cruz Biotechnology, Santa Cruz, CA, sc-66002, mouse monoclonal, 2 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, 20 minutes at 95 °C); anti-nestin (Santa Cruz, sc-23927, mouse monoclonal, 2 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, for 20 minutes at 95 °C, or DIVA for 30 minutes at 95 °C, or trypsin [DIFCO Laboratories, Detroit, MI, 0152-13] 1 mg/mL in 0.07 M CaCl 2 at pH 7.8 for 30 minutes at 37 °C); anti-S100 (Santa Cruz, sc-53438, mouse monoclonal, 0.8 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, 20 minutes at 95 °C); anti-class-III-β-tubulin (R&D Systems, Minneapolis, MN, MAB1195, mouse monoclonal, 2 μg protein/mL, 0.01 M citric acid-sodium citrate buffer, pH 6, 20 minutes at 95 °C); anti-phosphorylated neurofilament protein (Covance [now BioLegend] SMI-31 R, mouse monoclonal, ascites fluid, diluted 1:500, 0.01 M citric acid-sodium citrate buffer, pH 6, 20 minutes at 95 °C).

Techniques: Immunohistochemistry, Immunofluorescence