Journal: Scientific Reports

Article Title: METTL3-mediated m 6 A modification regulates OSCC progression via the HNRNPA2B1/FOXQ1 axis

doi: 10.1038/s41598-025-32025-7

Figure Lengend Snippet: METTL3 overexpression enhances proliferation, migration, invasion, and EMT in OSCC cells. The cell lines used in these assays underwent METTL3 overexpression or silencing transfection, with METTL3 expression verified by PCR and Western blot (Fig. A-B). ( A ) EdU assays in CAL27, oe-CAL27, and sh-CAL27 cell lines demonstrated that METTL3 overexpression significantly enhanced OSCC cell proliferation, while METTL3 silencing attenuated it. ( B , C ) Wound-healing and Transwell migration assays confirmed that METTL3 overexpression increased OSCC cell migration, whereas silencing reduced migration capacity. ( D ) Transwell invasion assays indicated that METTL3 overexpression enhanced invasive capacity, while silencing diminished it. ( E ) Analysis of EMT markers revealed increased E-cadherin and decreased N-cadherin in METTL3-silenced cells, with the opposite trend in METTL3-overexpressed cells, consistent with the METTL3/HNRNPA2B1/FOXQ1 axis promoting EMT through elevated FOXQ1 protein expression via HNRNPA2B1 binding to m 6 A-modified FOXQ1 mRNA. * p < 0.05, ** p < 0.01, *** p < 0.001 versus control.

Article Snippet: Next, the protein was separated by SDS-PAGE (P1200, Beijing Solabo Technology Co., Ltd.), transferred onto PVDF membranes (Millipore, USA), and cultivated with primary antibodies: METTL3 (1:1000, Rabbit, 15073-1-AP, Proteintech), HNRNPA2B1 (1:1000, Rabbit, ab31645, Abcam), FOXQ1 (1:500, Mouse, sc-166265, Santa Cruz), E-cadherin(1:1000, Rabbit, 20874-1-AP, Proteintech) and N-cadherin(1:1000, Rabbit, 22018-1-AP, Proteintech), followed by cultivation with secondary antibody.

Techniques: Over Expression, Migration, Transfection, Expressing, Western Blot, Binding Assay, Modification, Control

Journal: Scientific Reports

Article Title: METTL3-mediated m 6 A modification regulates OSCC progression via the HNRNPA2B1/FOXQ1 axis

doi: 10.1038/s41598-025-32025-7

Figure Lengend Snippet: Silencing METTL3 Reduces Tumorigenicity and FOXQ1 Expression in OSCC Xenografts. ( A ) Xenograft experiments were conducted in n = 6 male BALB/c nude mice per group (5-week-old, 20 ± 2 g) by injecting CAL27 or sh-CAL27 cells, with tumor growth monitored throughout the study. ( B ) Tumor weights were significantly higher in the CAL27 group compared to the sh-CAL27 group. ( C ) Tumor growth rates were higher in the CAL27 group compared to the sh-CAL27 group. ( D , E ) HE staining revealed keratinizing squamous epithelium and nuclear division in CAL27 tumors, features less pronounced in sh-CAL27 tumors; IHC staining showed significantly lower FOXQ1 expression in sh-CAL27 tumors compared to CAL27 tumors, with statistically significant differences. Additional IHC staining for EMT markers demonstrated elevated E-cadherin expression in sh-CAL27 tumors compared to CAL27 tumors, and reduced N-cadherin expression in sh-CAL27 tumors relative to CAL27 tumors, as quantified via ImageJ IHC Profiler. In vivo findings corroborate the METTL3/HNRNPA2B1/FOXQ1 axis, whereby METTL3-catalyzed m 6 A modification stabilizes HNRNPA2B1 mRNA and elevates its protein expression, facilitating HNRNPA2B1 binding to m 6 A-modified FOXQ1 mRNA to amplify FOXQ1 protein levels and drive tumor growth and EMT. * p < 0.05, ** p < 0.01, *** p < 0.001 versus control.

Article Snippet: Next, the protein was separated by SDS-PAGE (P1200, Beijing Solabo Technology Co., Ltd.), transferred onto PVDF membranes (Millipore, USA), and cultivated with primary antibodies: METTL3 (1:1000, Rabbit, 15073-1-AP, Proteintech), HNRNPA2B1 (1:1000, Rabbit, ab31645, Abcam), FOXQ1 (1:500, Mouse, sc-166265, Santa Cruz), E-cadherin(1:1000, Rabbit, 20874-1-AP, Proteintech) and N-cadherin(1:1000, Rabbit, 22018-1-AP, Proteintech), followed by cultivation with secondary antibody.

Techniques: Expressing, Staining, Immunohistochemistry, In Vivo, Modification, Binding Assay, Control

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