Journal: ACS biomaterials science & engineering

Article Title: Spatially Defined Cell-Secreted Protein Detection Using Granular Hydrogels: μ GeLISA

doi: 10.1021/acsbiomaterials.2c01308

Figure Lengend Snippet: (a) Representative maximum intensity projection of z-stack images of microgels incubated in different concentrations of MMP2. (i) 0, (ii) 300, (iii) 1200 ng/mL of MMP2. Purple: anti-MMP2. Green: microgel. Light blue: phalloidin 555. Scale bar: 50 μm (b) standard curve formed by measuring the average fluorescence of the microwells after being incubated at 0, 75, 150, 300, 600, and 1200 ng/mL. The measurements were then fit into a 4-parameter log-logistic regression. The curve has an inflection point at 457.28 ng/mL. Shaded area indicates 95% confidence interval. (c) Center z-slice of HSC spheroids with MMP2 μGeLISA. Purple: anti-MMP2. Light blue: phalloidin 555. Scale bar: 50 μm (d) average radial profile of MMP2 concentration around HSC aggregates of 10 and 50 cells. Shaded area shows the 95% confidence interval. n = 3 experimental replicates. (e) Total amount of secreted MMP2 by the aggregates within a 150 μm radius. Student’s t-test. *p < 0.05. n = 24–30 biological replicates.

Article Snippet: Recombinant human IL-6 protein (cat. no. 206-IL), human/primate IL-6 monoclonal antibody (dAB, cat. no. MAB206), biotinylated human/primate IL-6 (cAB, cat. no. BAF206), human MMP2 monoclonal antibody (MMP2 dAB, cat. no. MAB902), and human MMP2 biotinylated antibody (MMP2 cAB, cat. no. BAF902) were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Incubation, Fluorescence, Concentration Assay

Journal: Oncology Reports

Article Title: Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation

doi: 10.3892/or.2026.9086

Figure Lengend Snippet: Knockdown of CCT2 inhibits STAT3 signaling activation in hepatocellular carcinoma cells. The protein levels of STAT3, p-STAT3, MCL1, MMP2 and SOX2 in (A) Huh-7 and (B) HCCLM3 cells were measured by western blotting. The protein levels of (C) p-STAT3, (D) MCL1, (E) MMP2 and (F) SOX2 in subcutaneous tumor tissue were detected by immunohistochemical staining. *P<0.05, **P<0.01 vs. sh-NC. CCT2, chaperonin containing TCP1 subunit 2; sh, short hairpin; NC, negative control; p, phosphorylated; MCL1, myeloid cell leukemia sequence 1; SOX2, SRY-box transcription factor 2.

Article Snippet: The primary antibodies were as follows: CCT2 (cat. no. 24896-1-AP), β-actin (cat. no. 66009-1-Ig), MMP2 (cat. no. 10373-2-AP), myeloid cell leukemia sequence 1 (MCL1; cat. no. 16225-1-AP) and SRY-box transcription factor 2 (SOX2; cat. no. 11064-1-AP; all Proteintech Group, Inc.) and STAT3 (cat. no. 4904) and phosphorylated (p-)STAT3 (Tyr705; cat. no. 4113; both Cell Signaling Technology, Inc.) The membranes were washed three times in TBST (0.1% Tween-20) for 5 min each at room temperature.

Techniques: Knockdown, Activation Assay, Western Blot, Immunohistochemical staining, Staining, Negative Control, Sequencing