hmga2 3 utr reporter plasmids  (Addgene inc)

Journal: Frontiers in Pharmacology

Article Title: Effect of the antifungal drug ciclopirox on the inhibition of HMGA2-mediated oncogenic capacity in ACHN renal cell carcinoma

doi: 10.3389/fphar.2026.1723954

Figure Lengend Snippet: Effect of CPX on the expression of E-cadherin, N-cadherin, TGF-β R II, Smad-2, Smad-3, and Snail in tumor tissues. (A) Expression levels of E-cadherin, N-cadherin, TGF-β R II, Smad-2, Smad-3, and Snail in tumor tissues detected by qRT-PCR. (B) The expression levels of HMGA2, E2F1, Cyclin D1 and CDK6 in tumor tissues were examined by protein blotting (C) . Quantitation of the result of Western blot. Values are shown as mean ± SD. * P < 0.05 vs. control group, * ** P < 0.001 vs. control group.

Article Snippet: Antibodies against HMGA2 (ER62068), E2F1 (ET1701-73), CyclinD1 (ET1601-31), E-cadherin (ET1607-75), N-cadherin (ET1607-37), Smad2 (ET1604-22), Smad3 (ET1607-41), TGFR II (ER 1917-66), and Snai1 (ER1706-22) were purchased from HuaBio (Hangzhou, China), and CDK6 (ab124821) antibody was purchased from Abcam (Cambridge, United Kingdom).

Techniques: Expressing, Quantitative RT-PCR, Quantitation Assay, Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: Effect of the antifungal drug ciclopirox on the inhibition of HMGA2-mediated oncogenic capacity in ACHN renal cell carcinoma

doi: 10.3389/fphar.2026.1723954

Figure Lengend Snippet: Effect of CPX on the expression of HMGA2,E2F1, Cyclin D1, and CDK6 in renal cancer ACHN cells. (A) Expression levels of HMGA2,E2F1, Cyclin D1 and CDK6 in tumor tissues detected by qRT-PCR. (B) The expression levels of HMGA2, E2F1, Cyclin D1, and CDK6 in renal cancer ACHN cells examined by protein blotting. (C) Quantitation of the result of Western blot. Values are shown as mean ± SD. * P < 0.05 vs. control group, * ** P < 0.001 vs. control group. (D) The expression levels of HMGA2, E2F1, Cyclin D1, and CDK6 in renal cancer ACHN cells examined by qRT-PCR. (E) Quantitation of the result of Western blot.

Article Snippet: Antibodies against HMGA2 (ER62068), E2F1 (ET1701-73), CyclinD1 (ET1601-31), E-cadherin (ET1607-75), N-cadherin (ET1607-37), Smad2 (ET1604-22), Smad3 (ET1607-41), TGFR II (ER 1917-66), and Snai1 (ER1706-22) were purchased from HuaBio (Hangzhou, China), and CDK6 (ab124821) antibody was purchased from Abcam (Cambridge, United Kingdom).

Techniques: Expressing, Quantitative RT-PCR, Quantitation Assay, Western Blot, Control

Journal: Nature

Article Title: Critical role for a high-plasticity cell state in lung cancer

doi: 10.1038/s41586-025-09985-x

Figure Lengend Snippet: a , Representative H&E and immunohistochemistry (IHC) for Cre (to detect the Slc4a11-MCD reporter), uPAR, NKX2.1 and HMGA2 on serial sections from autochthonous KPfrt; Hipp11 GGCB/+ ; Slc4a11 FSF-MCD/+ LUAD tumours at 7 and 16 weeks PTI. Histopathological grades were assigned using Aiforia artificial intelligence (AI)-based image analysis software. Scale bar: 100 µm (low magnification) and 10 µm (high magnification). b , uPAR + ( top ), NKX2.1 + ( middle ), HMGA2 + ( bottom ) tumour cell percentages across histopathological grades. uPAR: n = 30 tumours per grade, 4 mice; NKX2.1: 28 tumours per grade, 4 mice; HMGA2: 31 tumours per grade, 4 mice. One-way ANOVA with Dunnett’s T3 multiple comparisons test. Error bars are SEM.

Article Snippet: The sections were blocked in donkey immunomix (0.2% BSA (Sigma-Aldrich, 810533), 5% donkey serum (Thermo Fisher Scientific, 31874), 0.3% Triton X-100 (Thermo Fisher Scientific, BP151-100) in PBS (Gibco, 10010-023)) at room temperature for 30 min. Incubation of primary antibodies against GFP (Abcam, ab13970), integrin α2 (Abcam, ab181548), Ki-67 (Thermo Fisher Scientific, 14-5698-82), pan-cytokeratin (Agilent Dako, M351529-2), uPAR (R&D Systems, AF807), Flag (Sigma-Aldrich, F1804), NKX2.1 (Abcam, ab76013), HMGA2 (Cell Signaling, 8179), SPC (Sigma-Aldrich, AB3786), HOPX (Santa Cruz Biotechnology, sc-398703), Cre recombinase (Cell Signaling, 15036) and HNF4α (Cell Signaling, 3113) diluted in donkey immunomix was performed at 4 °C overnight.

Techniques: Immunohistochemistry, Software

Journal: Nature

Article Title: Critical role for a high-plasticity cell state in lung cancer

doi: 10.1038/s41586-025-09985-x

Figure Lengend Snippet: a , Nkx2-1 ( top ) and Hmga2 ( bottom ) expression in LUAD cells from (Fig. ). b , IF showing mScarlet (white arrowheads) and NKX2.1 (blue arrowheads) mutual exclusivity in GFP + LUAD cells in KPfrt ; Hipp11 GGCB/+ ; Slc4a11 MCD/+ lung tumours at 16 weeks PTI. Scale bar: 20 µm. c , Percentage of NKX2.1 + cells in mScarlet − /GFP + vs. mScarlet + /GFP + LUAD cell subsets (n = 53 tumours, 5 mice; Mann-Whitney U test). d , IF showing mScarlet (white arrowheads) and HMGA2 (blue arrowheads) co-expression in subsets of GFP + LUAD cells in KPfrt ; Hipp11 GGCB/+ ; Slc4a11 MCD/+ lung tumours at 16 weeks PTI. Scale bar: 20 µm. e , Percentage of HMGA2 + cells in mScarlet − /GFP + vs. mScarlet + /GFP + subsets (n = 21 tumours, 5 mice; Mann-Whitney U test). f , Predicted terminal cell states (EMT, Proximal ciliated-like) and the HPCS projected on the LUAD progression PHATE map from Marjanovic*, Hofree*, Chan* et al. g , Expression of the indicated genes plotted as PHATE maps ( left ) and gene expression trajectories for the indicated marker genes plotted over Palantir pseudotime , with cell state trajectories predicted by CellRank , and estimated time spent in HPCS cell state shaded in red (see ‘ Time Series Analyses’ in ) ( right ).

Article Snippet: The sections were blocked in donkey immunomix (0.2% BSA (Sigma-Aldrich, 810533), 5% donkey serum (Thermo Fisher Scientific, 31874), 0.3% Triton X-100 (Thermo Fisher Scientific, BP151-100) in PBS (Gibco, 10010-023)) at room temperature for 30 min. Incubation of primary antibodies against GFP (Abcam, ab13970), integrin α2 (Abcam, ab181548), Ki-67 (Thermo Fisher Scientific, 14-5698-82), pan-cytokeratin (Agilent Dako, M351529-2), uPAR (R&D Systems, AF807), Flag (Sigma-Aldrich, F1804), NKX2.1 (Abcam, ab76013), HMGA2 (Cell Signaling, 8179), SPC (Sigma-Aldrich, AB3786), HOPX (Santa Cruz Biotechnology, sc-398703), Cre recombinase (Cell Signaling, 15036) and HNF4α (Cell Signaling, 3113) diluted in donkey immunomix was performed at 4 °C overnight.

Techniques: Expressing, MANN-WHITNEY, Gene Expression, Marker

Journal: Nature

Article Title: Critical role for a high-plasticity cell state in lung cancer

doi: 10.1038/s41586-025-09985-x

Figure Lengend Snippet: a-b , Quantification of HPCS in lineage-traced cells after tracing (n = 4 mice per time point; Mann-Whitney U test). c-d , mScarlet + /EpCAM + (HPCS/all cancer cells, c ) and GFP + /EpCAM + (Traced cells/all cancer cells, d ) at 3 days (3 d) or 14 days (14 d) of lineage tracing (6 to 8 weeks PTI). n = 4 mice per time point. Mann-Whitney U test. e-f , mScarlet + /EpCAM + (HPCS/all cancer cells, e ) and GFP + /EpCAM + (Traced cells/all cancer cells, f ) at 3 days (3 d) or 14 days (14 d) of lineage tracing (12 to 14 week). n = 4 mice per time point. Mann-Whitney U test. g , Cell states in combined tracing experiments from Fig. . scRNA-seq data from HPCS lineage-traced cells are plotted at the indicated timepoints. h , Stacked bar graphs showing the distribution of cell states across individual mice from HPCS tracing experiments shown as stacked bar graphs as in Fig. . Error bars are SEM. i , Distribution of Nkx2-1 , Sftpc , Hopx , and Hmga2 expression from traced LUAD cells at 8 weeks (6 to 8 week tracing, left ) or 14 weeks (12 to 14 week tracing, right ) PTI. j , Percentage of HPCS (red) or other cell states (green) expressing the indicated genes in traced LUAD cells at 8 weeks (6 to 8 week tracing, left ) or 14 weeks (12 to 14 week tracing, right ) PTI. Fisher’s exact test. k , IF images showing co-staining of GFP (green) with either NKX2.1 ( left , red), SPC ( middle , red), or HMGA2 ( right , red). Boxed areas are shown as individual insets below their respective images. Scale bar: 20 µm. l , Percentage of NKX2.1 + ( left ), SPC + ( middle ) or HMGA2 + ( right ) cells in GFP + LUAD cells quantified from IF co-staining of LUAD tissues harvested at 14 weeks PTI after 3 (3 d) and 14 (14 d) days post-tracing. NKX2.1: n = 21 tumours, 3 mice (3 d) and n = 111 tumours, 3 mice (14 d). SPC: n = 59 tumours, 3 mice (3 d) and n = 89 tumours, 3 mice (14 d). HMGA2: n = 84 tumours, 3 mice (3 d) and n = 35 tumours, 3 mice (14 d). Mann-Whitney U test. Error bars are SEM.

Article Snippet: The sections were blocked in donkey immunomix (0.2% BSA (Sigma-Aldrich, 810533), 5% donkey serum (Thermo Fisher Scientific, 31874), 0.3% Triton X-100 (Thermo Fisher Scientific, BP151-100) in PBS (Gibco, 10010-023)) at room temperature for 30 min. Incubation of primary antibodies against GFP (Abcam, ab13970), integrin α2 (Abcam, ab181548), Ki-67 (Thermo Fisher Scientific, 14-5698-82), pan-cytokeratin (Agilent Dako, M351529-2), uPAR (R&D Systems, AF807), Flag (Sigma-Aldrich, F1804), NKX2.1 (Abcam, ab76013), HMGA2 (Cell Signaling, 8179), SPC (Sigma-Aldrich, AB3786), HOPX (Santa Cruz Biotechnology, sc-398703), Cre recombinase (Cell Signaling, 15036) and HNF4α (Cell Signaling, 3113) diluted in donkey immunomix was performed at 4 °C overnight.

Techniques: MANN-WHITNEY, Expressing, Staining