Journal: BMC Cancer

Article Title: Elevated expression of CXCL3 in colon cancer promotes malignant behaviors of tumor cells in an ERK-dependent manner

doi: 10.1186/s12885-023-11655-y

Figure Lengend Snippet: CXCL3 overexpression modulates ERK pathway-related protein expression in COAD cells. ( A, C ) Representative images of western blot assay for protein expression. ( B, D ) Analysis of protein levels (normalized to β-actin). * P < 0.05 vs. Mock, ** P < 0.01 vs. Mock

Article Snippet: Following blocking with 5% skimmed milk, the membranes were treated with various specific antibodies, including a rabbit antibody against ERK1/2 (1:1,000 dilution; Abways Technology Company), a rabbit antibody against p-ERK1/2 (1:1,000 dilution; Abways Technology Company), a rabbit antibody against Bcl-2 (1:1,000 dilution; Abways Technology Company), a rabbit antibody against Bax (1:1,000; Abways Technology Company), a rabbit antibody against Cyclin D1 (1:1,000; Abways Technology Company) and a mouse antibody against β-actin (1:1,000; Zhongshan Golden Bridge Bio-technology, Beijing).

Techniques: Over Expression, Expressing, Western Blot

Journal: Nature Communications

Article Title: A reversible state of hypometabolism in a human cellular model of sporadic Parkinson’s disease

doi: 10.1038/s41467-023-42862-7

Figure Lengend Snippet: a Quantification of total mitochondrial mass and functional mitochondria in hNPCs or b DAns. Total mitochondrial mass was visualized using immunostainings with an antibody against the ATP synthase (ATP5F1A). Functional mitochondria with an active membrane potential were visualized using a MitoTracker probe (200 nM for 20 min). Images are exemplarily shown for hNPCs of clone i1E4-R1-003 (Ctrl), and iR66-R1-007 (sPD); for DAns of clone i1E4-R1-003 (mitoTracker—Ctrl), iR66-R1-007 (mitoTracker—sPD), i1JF-R1-018 (ATP5F1A—Ctrl), and iJ2C-R1-015 (ATP5F1A—sPD). Scale bar = 20 μm. c Violin plots highlight some morphological characteristics of mitochondria in hNPCs or d DAns: number of mitochondria (count), mitochondrial area, skeleton length, number of branch points, and mean intensity. On average, 112 (hNPCs—ATP5F1A), 160 (hNPCs—mitoTracker), 370 (DAns—ATP5F1A), 360 (DAns—mitoTracker) cells per clone were analyzed. e To assess alterations in the mitochondrial fusion machinery, expression of the mitofusions MFN1 , MFN2 , and OPA1 was quantified in hNPCs by RT-qPCR. Cells were cultivated on the energy substrates used in the Seahorse XF analysis (25 mM glucose or 5 mM pyruvate). f To assess alterations in the mitochondrial fission machinery, expression of DRP1 and its phosphorylation on Ser 616 was quantified in hNPCs by western blot. Protein levels were normalized to ACTB. g Western blots are exemplary shown for some hNPC clones. Boxplots display the median and range from the 25th to 75th percentile. Whiskers extend from the min to max value or to the most extreme data point which is no more than 1.5 times the interquartile range ( c , d ). Each dot represents one patient. n = 5 Ctrl and 7 sPD patient-derived cell clones, in triplicates. p- values were determined by linear mixed effects model ( c , d ); one-way ANOVA with Sidak’s Post hoc test ( p -values are provided together with the source data) ( e , f ). * p < 0.05; ** p < 0.01; *** p < 0.001. Source data are provided as a Source Data file.

Article Snippet: Primary antibodies NDUFB8 (459210, Novex; 1:500), Complex II- Subunit 30 (SDHB) (459230, Thermo Fisher Scientific; 1:500), UQCRC2 (ab14745, Abcam; 1:2500), MT-CO2 (ab110258, Abcam; 1:1000), ATP5F1A (ab14748, Abcam; 1:4000), DRP1 (5391, Cell Signaling; 1:1000), DRP1 phospho-Ser616 (4494, Cell Signaling; 1:1000), TUBA (GTX628802, Genetex; 1:20,000), ACTB (ABO145-200, OriGene; 1:2000), OGDHL (17110-1-AP, Proteintech, 1:5000), GAPDH (GTX627408 peroxidase coupled, Genetex, 1:20,000), SLC2A1 (MA5-31960, Invitrogen, 1:5000), SLC2A3 (PA5-72331, Thermo Fisher Scientific, 1:5000) were incubated in blocking buffer overnight at 4 °C.

Techniques: Functional Assay, Membrane, Expressing, Quantitative RT-PCR, Phospho-proteomics, Western Blot, Clone Assay, Derivative Assay