Journal: The Journal of Pathology: Clinical Research

Article Title: The prognostic significance of the IASLC grading system in ALK ‐positive invasive non‐mucinous adenocarcinoma of the lung: correlation with clinicopathological features

doi: 10.1002/2056-4538.70093

Figure Lengend Snippet: Histological and immunohistochemical features of the ALK‐positive INMA cases (A) Examples of weak or strong ALK positivity (middle and right), with the left panel demonstrating negative staining. (B) H&E‐stained sections demonstrating histological patterns: lepidic (left), acinar (middle), and papillary (right). (C) Representative H&E‐stained sections showing cribriform (left), micropapillary (middle), and complex glandular (right) patterns. (D) H&E staining of an example of solid growth pattern (left); IHC staining shows tumor cells positive for TTF‐1 (middle) and negative for p40 (right). (bar = 100 μm). ALK, anaplastic lymphoma kinase; H&E, hematoxylin and eosin; IHC, immunohistochemistry; INMA, invasive non‐mucinous adenocarcinoma.

Article Snippet: The antibodies used included: ALK (790‐4797, ready‐to‐use, Ventana®), TTF‐1 (11833669A, 1:100, Zhongshan Golden Bridge Biotechnology, Beijing, China), p40 (25081519, 1:100, Dako, Carpinteria, CA, USA), Ki‐67 ( GT209429 , 1:100, Gene Tech Company Limited, Shanghai, China), PD‐L1 (SK006, ready‐to‐use, Dako, Carpinteria, CA, USA), CD4 (IR649, 1:100, Dako, Carpinteria, CA, USA), CD8 (ZA‐0508, 1:100, Zhongshan Golden Bridge Biotechnology, Beijing, China), and FoxP3 (sc‐53876, 1:100, Santa Cruz, Dallas, TX, USA).

Techniques: Immunohistochemical staining, Negative Staining, Staining, Immunohistochemistry

Journal: Cell reports

Article Title: Pharmacological administration of FGF21 reverses obesity through a parabrachial-projecting neuron population in the hindbrain

doi: 10.1016/j.celrep.2026.117093

Figure Lengend Snippet: (A–D) Diet-induced obese (DIO) WT and KLB Nkx2.1–Cre (KLB fl/fl ; Nkx2.1-Cre) mice (16–18 weeks old) were administered vehicle or FGF21 (1 mg/kg/day) by osmotic minipump for 14 days ( n = 9–13 per group). (A) Plasma FGF21 levels, (B) starting body weights, (C) daily percent change in body weight, and (D) final percent change in body weight. (E and F) DIO WT and KLB NKX2.1–Cre mice (16–18 weeks old) were administered vehicle or PF-05231023 (i.p. 3 mg/kg) every 3 days for 15 days ( n = 8–11 per group). (E) Daily percent change in body weight and (F) final percent change in body weight. (G–J) DIO WT and KLB loxTB (KLB loxTB/loxTB ) mice (16–18 weeks old) were injected with AAV-CMV-Cre-EGFP (KLB loxTB ; Cre) or AAV-CMV-eGFP (KLB loxTB ; GFP) in the hypothalamus and administered vehicle or FGF21 (1 mg/kg/day) by osmotic minipump for 14 days ( n = 6–13 per group). (G) Schematic illustration of the viral targeting strategy to restore endogenous KLB expression specifically in the hypothalamus, (H) plasma FGF21 levels, (I) daily percent change in body weight, and (J) final percent change in body weight of mice in (G)–(J). Values are presented as mean ± SEM. * p < 0.05. Statistical analyses were conducted using Student’s t test (B) or two-way ANOVA (C–F, I, and J) with Holms-Sidak’s multiple comparison test.

Article Snippet: Nkx2.1-Cre , The Jackson Laboratory , 008661.

Techniques: Clinical Proteomics, Injection, Expressing, Comparison

Journal: International Journal of Biological Sciences

Article Title: NOX4-derived oxidative DNA damage impairs thyroid differentiation through an epigenetic mechanism in BRAF-mutated radioactive iodine refractory papillary thyroid cancer cells

doi: 10.7150/ijbs.123980

Figure Lengend Snippet: DNMT inhibition and knockdown of MSH2, MSH6, and OGG1 synergize to increase NIS mRNA expression. A) qRT-PCR analysis of NIS mRNA levels in BCPAP cells transfected with siRNA control or/and siRNA MSH2 or/and siRNA MSH6 or/and siRNA OGG1 and 24 h later treated for an additional 48 h in the presence or the absence of 1 µM DAC. B) qRT-PCR analysis of NIS mRNA levels in BCPAP cells transfected with siRNA control or siRNA NOX4 or siRNA p22 phox and 24 h later, treated for an additional 48 h in the presence or the absence of 1 µM DAC. Values are mean ± SE. *p < 0.05, **p < 0.01, and ***p < 0.001. C) Western blot analysis of PAX8 and NKX2.1 protein expressions in chromatin fraction and whole-cell extract 72 h after knocking down of NOX4 or p22 phox by RNA interference in BCPAP cells. Densitometry quantification of protein levels normalized to Lamin B or Vinculin levels and presented as chromatin enrichment or fold change compared with control cells. Values are mean ± SE. *p < 0.05 and **p < 0.01 (n = 5). D) ChIP-qPCR assays performed with BCPAP cells transfected with siRNA control and with BCPAP cells transfected with siRNA NOX4 immunoprecipitated with control IgG or anti-NKX2.1 antibody and analyzed by qPCR at TG promoter (two independent replicates).

Article Snippet: Subsequently, the chromatin fragments were immunoprecipitated with 1 μg of anti-NKX2.1 (TTF1) antibody (Cell Signaling Technology, #12373, RRID: AB_2797895) or an equal amount of rabbit IgG isotype control (Agilent, #0903).

Techniques: Inhibition, Knockdown, Expressing, Quantitative RT-PCR, Transfection, Control, Western Blot, ChIP-qPCR, Immunoprecipitation

Journal: International Journal of Biological Sciences

Article Title: NOX4-derived oxidative DNA damage impairs thyroid differentiation through an epigenetic mechanism in BRAF-mutated radioactive iodine refractory papillary thyroid cancer cells

doi: 10.7150/ijbs.123980

Figure Lengend Snippet: OGG1, MSH2, and MSH6 inhibit recruitment of PAX8 and NKX2.1 to chromatin and the MAPK pathway regulates MSH2 and MSH6 expressions. A) Western blot analysis of PAX8 and NKX2.1 protein expressions in chromatin fraction and whole-cell extract 72 h after knocking down of OGG1 (siRNA OGG1#1), MSH2, or/and MSH6 by RNA interference in BCPAP cells. Densitometry quantification of protein levels normalized to Lamin B or Vinculin levels and presented as chromatin enrichment and fold change compared with control cells. Values are mean ± SE. *p < 0.05, **p < 0.01, and ***p < 0.001 (n = 3). B) Immunoblot detection of MSH2 and MSH6 in PDX cells after treatment by the combination of dabrafenib (100 nM) plus trametinib (5 nM). Densitometric quantification of protein levels normalized to vinculin levels and presented as fold change compared with vehicle-treated cells. Student t-test is realized by comparing combination versus DMSO for each corresponding time of kinetic. Values are mean ± SE. *p < 0.05 and **p < 0.01 (n = 3).

Article Snippet: Subsequently, the chromatin fragments were immunoprecipitated with 1 μg of anti-NKX2.1 (TTF1) antibody (Cell Signaling Technology, #12373, RRID: AB_2797895) or an equal amount of rabbit IgG isotype control (Agilent, #0903).

Techniques: Western Blot, Control

Journal: International Journal of Biological Sciences

Article Title: NOX4-derived oxidative DNA damage impairs thyroid differentiation through an epigenetic mechanism in BRAF-mutated radioactive iodine refractory papillary thyroid cancer cells

doi: 10.7150/ijbs.123980

Figure Lengend Snippet: MAPK pathway inhibition and knockdown of NOX4 and p22 phox synergize to promote PAX8 and NKX2.1 recruitment to chromatin. A) Western-blot analysis of PAX8 and NKX2.1 protein expressions in chromatin fractions and whole-cell extracts of BCPAP cells transduced with siRNA control or siRNA NOX4 or siRNA p22 phox and treated with dabrafenib (100 nM) plus trametinib (25 nM) combination for 48 h. Densitometric quantification of protein levels normalized to Lamin B or loading control (red ponceau staining) and presented as chromatin enrichment and fold change compared with siRNA control-transduced cells. B) qRT-PCR analysis of NIS mRNA levels in BCPAP cells transfected with siRNA control or siRNA NOX4 or siRNA p22 phox and 24 h later, treated in the presence or the absence of dabrafenib plus trametinib combination for additional 48 h. C) qRT-PCR analysis of TSHR mRNA levels in BCPAP cells transfected with siRNA control or siRNA NOX4 or siRNA p22 phox and 24 h later, treated in the presence or the absence of dabrafenib plus trametinib combination for additional 48 h. Values are mean ± SE. *p < 0.05, **p < 0.01, and ***p < 0.001 (n = 3).

Article Snippet: Subsequently, the chromatin fragments were immunoprecipitated with 1 μg of anti-NKX2.1 (TTF1) antibody (Cell Signaling Technology, #12373, RRID: AB_2797895) or an equal amount of rabbit IgG isotype control (Agilent, #0903).

Techniques: Inhibition, Knockdown, Western Blot, Transduction, Control, Staining, Quantitative RT-PCR, Transfection

Journal: International Journal of Biological Sciences

Article Title: NOX4-derived oxidative DNA damage impairs thyroid differentiation through an epigenetic mechanism in BRAF-mutated radioactive iodine refractory papillary thyroid cancer cells

doi: 10.7150/ijbs.123980

Figure Lengend Snippet: Co-inhibition of SMAD and MAPK signalling promotes PAX8 and NKX2.1 recruitment to chromatin. A) BRAF-mutated thyroid cells were treated with TGF-beta receptor inhibitor (EW7197, 1 µM) for 4 h and analysed by Western-blot for expression of pSMAD3 and SMAD3. B) Western-blot analysis of PAX8 and NKX2.1 protein expressions in chromatin fractions and whole-cell extracts of BCPAP cells treated or not with dabrafenib (100 nM) plus trametinib (25 nM) combination in the presence or the absence of TGF-beta inhibitor (EW7197, 1 µM) for 48 h. Densitometric quantification of protein levels normalized to loading control and presented as chromatin enrichment or fold change compared with vehicle-treated cells. C and D) qRT-PCR analysis of NIS and TSHR mRNA levels in BCPAP cells treated or not with dabrafenib plus trametinib combination in the presence or the absence of TGF-beta inhibitor (EW7197) for 48 h. Densitometry quantification of protein levels normalized to Lamin B or Vinculin levels and presented as fold change compared with control cells. Values are mean ± SE. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p<0.0001 (n = 3).

Article Snippet: Subsequently, the chromatin fragments were immunoprecipitated with 1 μg of anti-NKX2.1 (TTF1) antibody (Cell Signaling Technology, #12373, RRID: AB_2797895) or an equal amount of rabbit IgG isotype control (Agilent, #0903).

Techniques: Inhibition, Western Blot, Expressing, Control, Quantitative RT-PCR