Journal: The Journal of international medical research

Article Title: MYL9 promotes squamous cervical cancer migration and invasion by enhancing aerobic glycolysis.

doi: 10.1177/03000605231208582

Figure Lengend Snippet: Figure 3. MYL9 promoted the progression of SCC by enhancing aerobic glycolysis. (a) MYL9 knockdown decreased the Lactate production of SCC. (b) MYL9 knockdown inhibited the expression of GLUT1, HK2, and LDHA and (c) MYL9 knockdown inactivated the JAK2/STAT3 pathway. MYL9, myosin light chain 9; SCC, squamous cervical cancer; GLUT1, glucose transporter 1; HK2, hexokinase II; LDHA, lactate dehydrogenase A.

Article Snippet: The membranes were blocked with 5% bovine serum albumin in TBS containing Tween-20 (TBST) for 1 hour at 37 C and incubated with primary antibodies againstMYL9 (mouse, 60233-1-Ig, Proteintech, Rosemont, IL, USA), GLUT1 (rabbit, 21829-1-AP, Proteintech), HK2 (mouse, 66974-1-Ig, Proteintech), LDHA (rabbit, 19987-1-AP, Proteintech), b-tubulin (mouse, Proteintech), and GAPDH (mouse, CW0100M, CWBIO, Beijing, China) overnight at 4 C. Following three washes with TBST, the membrane was incubated with horseradish peroxidase-conjugated goat antirabbit or anti-mouse IgG antibody at 37 C for 1 hour.

Techniques: Knockdown, Expressing

Journal: Molecular metabolism

Article Title: Increased α-HB links colorectal cancer and diabetes by potentiating NF-κB signaling.

doi: 10.1016/j.molmet.2023.101766

Figure Lengend Snippet: Figure 5: a-HB-induced LDHA nuclear translocation extends NF-kB nuclear retention. (A) Cellular distribution of LDHA in SW620 (upper) and MIN6 (bottom) cells treated with different concentrations of a-HB for 48 h. (BeC) ICC images of LDHA (green), p65 (red, upper) and p50 (red, bottom) in SW620 cells after 48 h-preincubation of a-HB and/or LPS treatment for 1.5 h (B) and 12 h (C), as indicated by DAPI (blue). Scale bars, 7.5 mm. (D) Nuclear LDHA accumulation over time in SW620 and MIN6 cells with continuous a-HB and/ or LPS treatment. (E) The time course of LDHA nuclear retention in SW620 and MIN6 cells after 48 h of a-HB treatment. (F) Immunoprecipitation analysis of the interactions among LDHA (upper), p65 (middle) and p50 (bottom) in SW620 cells treated with a-HB for 48 h and/or LPS for 1.5 h. (G) The effects of p65 phosphorylation sites on the interaction between LDHA and p65 were detected in p65 WT/mutant plasmids tranfected-SW620 cells treated with a 48 h-preincubated a-HB treatment and/or LPS stimulation for 1.5 h.

Article Snippet: Antibodies and reagents Antibodies against LDHA (Cell Signaling Technology, 3582, 1:300 for ICC and IHC; Santa Cruz, 1:50 working dilution for co-IP; Proteintech, 19987-1-AP, 1:2000 working dilution for western blotting), p65 (Cell Signaling Technology, 8242, 1:1000 working dilution for western blotting, 1:300 for ICC; Cell Signaling Technology, 6956, 1:100 for coIP), p50 (Cell Signaling Technology, 13,586, 1:1000 working dilution for western blotting, 1:300 for ICC, Santa Cruz, sc-8414, 1: 50 for coIP), phospho-p65 (Cell Signaling Technology, 3033, 1:1000 working dilution for western blotting), FOXM1 (Proteintech, 13147-1-AP, 1:2000 for western blotting), c-MYC (Proteintech, 10828-1-AP, 1:2000 for western blotting), a-tubulin (Proteintech, HRP-66031, 1:10,000 working dilution for western blotting), Histone H3 (Cell Signaling Technology, 4499, 1:1000 working dilution for western blotting), Lamin B (Santa Cruz, sc-6217, 1:500 working dilution for western blotting), b-actin (Santa Cruz, sc-47778, 1:500 working dilution for western blotting), HA (MBL, 1:100 working dilution for western blotting), CD4 (Abcam, ab183685, 1:1000 working dilution for IHC), and CD8 (Abcam, ab217344, 1:1000 working dilution for IHC) were purchased commercially.

Techniques: Translocation Assay, Immunoprecipitation, Phospho-proteomics, Mutagenesis

Journal: Cancer Science

Article Title: The Warburg effect as a therapeutic target for bladder cancers and intratumoral heterogeneity in associated molecular targets

doi: 10.1111/cas.15047

Figure Lengend Snippet: mRNA expression of putative targets associated with the Warburg effect in Ta, T1, and muscle‐invasive bladder cancer (MIBC) clinical specimens. mRNA expression analysis (log2) of the indicated targets associated with the Warburg effect (LDH‐A, GLUT1, MCT4) and intratumoral metabolic coupling (lactate import transporter MCT1) in 263 urothelial tumor specimens (125 Ta G2, 106 T1 G3, and 32 MIBC clinical specimens). LDH‐A/LDH‐B indicates ratio of LDH‐A expression relative to LDH‐B. Box and whisker plots showing the median, mean ( x ), and minimum and maximum expression following clustering by stage. * P < .05, ** P < .01; Student's t ‐test

Article Snippet: Antibodies used were: GLUT1 (ab15309, Abcam), LDH‐A (19987‐1‐AP, Proteintech), MCT4 (H90, Santa Cruz Biotechnology), and MCT1 (AB3538P, Merck Millipore).

Techniques: Expressing, Whisker Assay