Journal: Nature Communications

Article Title: Colonic epithelial-derived FGF1 drives intestinal stem cell commitment toward goblet cells to suppress inflammatory bowel disease

doi: 10.1038/s41467-025-58644-2

Figure Lengend Snippet: A Heatmap of differentially expressed Atoh1 and its downstream target genes in PBS and rFGF1-treated UC mice ( n = 4). The mRNA level of Atoh1 in PBS or rFGF1-treated UC mice (upper panel, n = 4) and CD mice (lower panel, n = 5) was detected by qRT-PCR. B , C IF staining of ATOH1 in distal colon sections of Fgf1 fl/fl and VilCre Fgf1 fl/fl mice challenged with DSS ( B ) or TNBS ( C ), and its semi-quantitation of IF intensity ( n = 4, the mean value of 2 fields in each mouse). D , E IF staining of ATOH1 in distal colon sections of DSS ( D ) or TNBS ( E )-induced two IBD mouse models, followed by PBS or rFGF1 treatment and its semi-quantitation of IF intensity ( n = 4, the mean value of 2 fields in each mouse). F Representative images (upper panel) and IF staining of Muc2 (lower panel) in sh Atoh1 or shNC-transfected colonic organoids stimulated with vehicle or rFGF1 ( n = 6). G , H Deletion of Atoh1 in VilCre ERT2 Atoh1 fl/fl mice was confirmed by qRT-PCR ( G ) and IF staining of colon tissues ( H ) ( n = 4). I – M VilCre ERT2 Atoh1 fl/fl mice were injected with tamoxifen for five consecutive days and then given drinking water containing 1.5% DSS to induce acute colitis, followed by vehicle or rFGF1 administration for 7 days. At end of the experiment, the distal colon tissues were harvested and examined. The schematic diagram shows the strategy of tamoxifen injection, DSS challenge and rFGF1 administration ( n = 4). J – L Weight loss ( J ), disease activity index ( K ), colonic length ( L ) were monitored (n = 4). M H&E (upper panel), PAS-AB staining (middle panel) and Muc2 IF staining (lower panel) of distal colon sections ( n = 4). Data was presented as mean ± SEM. ( A , D – E , H ) two-tailed unpaired t -test; ( B , C , F , J – M ) ordinary two-way ANOVA, followed by Sidak; ( G ) Non-parametric statistical method, two-tailed Mann-Whitney test. ns, not significance; nd, not detectable.

Article Snippet: The primary antibodies are listed below: rabbit anti-FGF1 (1:200, 17400, Proteintech), mouse anti-FGF1 (1:100, sc-55520, Santa Cruz Biotechnology), EpCAM (1:800, CST42515, Cell Signaling Technology), Col1α1 (1:100, CST72026, Cell Signaling Technology), ATOH1 (1:400, 21215-1-AP, Proteintech), TCF4 (1:100, ab217668, Abcam), Muc2 (1:400, ab272692, Abcam), Dclk1 (1:300, CST62257, Cell Signaling Technology), ChgA (1:200, sc-393941, Santa Cruz Biotechnology), Fabp1 (1:50, CST13368, Cell Signaling Technology), Ki67 (1:400, ab15580, Abcam), FGFR2 (1:200, CST23328S, Cell Signaling Technology) and Lgr5 (1:100, UM870104, ORIGENE).

Techniques: Quantitative RT-PCR, Staining, Quantitation Assay, Transfection, Injection, Activity Assay, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: Colonic epithelial-derived FGF1 drives intestinal stem cell commitment toward goblet cells to suppress inflammatory bowel disease

doi: 10.1038/s41467-025-58644-2

Figure Lengend Snippet: A Workflow to find the rFGF1-regulated potential transcription factors of Atoh1 , and heatmap of expression level of potential transcription factors ( n = 4). B The expression levels of potential transcription factors of Atoh1 ( Tcf4 , Nr4a1 , Nfya , Tec , Hoxa13 , Ncaph2 , Kdm1a ) in distal colon sections of PBS or rFGF1-treated UC mice were analyzed by qRT-PCR ( n = 4). C , D IF staining of TCF4 in distal colon sections of Fgf1 fl/fl and VilCre Fgf1 fl/fl mice challenged with DSS ( C ) or TNBS ( D ) and its semi-quantitation of IF intensity ( n = 4, the mean value of 2 fields in each mouse). E , F The expression levels of TCF4 in distal colon sections of DSS-treated UC mice ( E ) and TNBS-treated CD mice ( F ) after PBS or rFGF1 administration and IF staining ( n = 8). G Predicted TCF4 binding sites on the promoter of Atoh1 by the JASPAR database (left panel). Six binding sites with the most significant prediction scores were selected for pairwise deletion mutation analysis (right panel; M1, in purple; M2, in green; M3, in red). H , I Luciferase activity in HT-29 cells transfected with a human Atoh1 promoter reporter plasmid or the predicted TCF4 binding site mutant reporter along with a Tcf4 overexpression plasmid ( n = 4). J ChIP experiments. The data were normalized to the corresponding IgG ( n = 4). K Images of murine colonic organoids transfected small hairpin RNA targeting Tcf4 (sh Tcf4 ) or negative control hairpin (shNT) followed by treated with rFGF1 or vehicle ( n = 6). L IF staining of ATOH1 (upper panel) and Muc2 (lower panel) in colonic organoids in different groups and its semi-quantitation of fluorescent intensity ( n = 6). Data was presented as mean ± SEM. ( B , right panel of E – F , J ) two-tailed unpaired t -test ( Tec and Ncaph2 in penal B using non-parametric statistical method, two-tailed Mann-Whitney Test); ( H – I ) ordinary one-way ANOVA, followed by Dunnett; (right panel of C – D and L , lower panel of K ) ordinary two-way ANOVA, followed by Sidak. ns, no significance.

Article Snippet: The primary antibodies are listed below: rabbit anti-FGF1 (1:200, 17400, Proteintech), mouse anti-FGF1 (1:100, sc-55520, Santa Cruz Biotechnology), EpCAM (1:800, CST42515, Cell Signaling Technology), Col1α1 (1:100, CST72026, Cell Signaling Technology), ATOH1 (1:400, 21215-1-AP, Proteintech), TCF4 (1:100, ab217668, Abcam), Muc2 (1:400, ab272692, Abcam), Dclk1 (1:300, CST62257, Cell Signaling Technology), ChgA (1:200, sc-393941, Santa Cruz Biotechnology), Fabp1 (1:50, CST13368, Cell Signaling Technology), Ki67 (1:400, ab15580, Abcam), FGFR2 (1:200, CST23328S, Cell Signaling Technology) and Lgr5 (1:100, UM870104, ORIGENE).

Techniques: Expressing, Quantitative RT-PCR, Staining, Quantitation Assay, Binding Assay, Mutagenesis, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Over Expression, Negative Control, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: Colonic epithelial-derived FGF1 drives intestinal stem cell commitment toward goblet cells to suppress inflammatory bowel disease

doi: 10.1038/s41467-025-58644-2

Figure Lengend Snippet: A Single-cell RNA sequencing analysis (GSM4983265) showed that Fgfr2 is highly expressed in stem cells. B Representative image of colon distal stained with Lgr5 (green), FGFR2 (red) and DAPI (blue) from the C57BL/6 J mice ( n = 3). C Deletion of Fgfr2 in Lgr5-EGFP-Cre ERT2 Fgfr2 fl/fl mice treated with vehicle or tamoxifen was confirmed by IF staining ( n = 5). D Murine colonic organoids derived from vehicle or tamoxifen-treated Lgr5-EGFP-Cre ERT2 Fgfr2 fl/fl mice were stimulated with vehicle or rFGF1. Representative images, IF staining of TCF4, ATOH1 and Muc2 in colonic organoids were shown (n = 6). E – I Vehicle or tamoxifen-treated Lgr5-EGFP-Cre ERT2 Fgfr2 fl/fl mice were challenged with drinking water containing 2.5% DSS to induce UC, followed by colon tissues were harvested and examined (n = 4 for TAM + PBS group, n = 5 for other groups). Weight loss ( E ), disease activity index ( F ) and colon length ( G ) of different groups were monitored. H H&E staining of distal colon sections and histology scores (upper panel), PAS-AB staining of distal colon sections and mucin granules-positive cells per crypt (lower panel). I IF staining of TCF4 (upper panel), ATOH1 (middle panel) and Muc2 (lower panel) in distal colon sections, and its semi-quantification of IF intensity. Data was presented as mean ± SEM. (right panel of C ) two-tailed unpaired t -test; (right panel of D – F , right panel of G – I ) ordinary two-way ANOVA, followed by Sidak. ns, no significance.

Article Snippet: The primary antibodies are listed below: rabbit anti-FGF1 (1:200, 17400, Proteintech), mouse anti-FGF1 (1:100, sc-55520, Santa Cruz Biotechnology), EpCAM (1:800, CST42515, Cell Signaling Technology), Col1α1 (1:100, CST72026, Cell Signaling Technology), ATOH1 (1:400, 21215-1-AP, Proteintech), TCF4 (1:100, ab217668, Abcam), Muc2 (1:400, ab272692, Abcam), Dclk1 (1:300, CST62257, Cell Signaling Technology), ChgA (1:200, sc-393941, Santa Cruz Biotechnology), Fabp1 (1:50, CST13368, Cell Signaling Technology), Ki67 (1:400, ab15580, Abcam), FGFR2 (1:200, CST23328S, Cell Signaling Technology) and Lgr5 (1:100, UM870104, ORIGENE).

Techniques: RNA Sequencing, Staining, Derivative Assay, Activity Assay, Two Tailed Test

Journal: Molecular Medicine Reports

Article Title: Effect of Hath1 on the proliferation and apoptosis of cutaneous squamous cell carcinoma in vitro

doi: 10.3892/mmr.2015.4463

Figure Lengend Snippet: Relative mRNA expression of Hath1 in KUMA5 cells. Control, blank control group (Dulbecco's modified Eagle's medium/F12); GFP, pGen-esil-1 scramble control group; Hath1, pcDNA3.1-Hath1-transfected group; shRNA, pGenesil-1-Hath1-transfected group. GFP, green fluorescent protein; shRNA, short hairpin RNA.

Article Snippet: The PVDF membranes were then blocked in Tris-buffered saline (TBS; 10 mM Tris-HCl, pH 7.5 and 150 mM NaCl) containing 5% non-fat dry milk at room temperature for 1 h and incubated at 4° C overnight with anti-Hath1 monoclonal antibody (1:400; cat. no. sc-136173; Santa Cruz Biotechnology, Inc.) and anti-β-actin monoclonal antibody (1:1,000; cat. no. sc-47778; Santa Cruz Biotechnology, Inc.).

Techniques: Expressing, Control, Modification, Transfection, shRNA

Journal: Molecular Medicine Reports

Article Title: Effect of Hath1 on the proliferation and apoptosis of cutaneous squamous cell carcinoma in vitro

doi: 10.3892/mmr.2015.4463

Figure Lengend Snippet: Relative protein expression of Hath1 in KUMA5 cells. Control, blank control group (Dulbecco's modified Eagle's medium/F12); GFP, pGen-esil-1 scramble control group; Hath1, pcDNA3.1-Hath1-transfected group; shRNA, pGenesil-1-Hath1-transfected group. GFP, green fluorescent protein; shRNA, short hairpin RNA.

Article Snippet: The PVDF membranes were then blocked in Tris-buffered saline (TBS; 10 mM Tris-HCl, pH 7.5 and 150 mM NaCl) containing 5% non-fat dry milk at room temperature for 1 h and incubated at 4° C overnight with anti-Hath1 monoclonal antibody (1:400; cat. no. sc-136173; Santa Cruz Biotechnology, Inc.) and anti-β-actin monoclonal antibody (1:1,000; cat. no. sc-47778; Santa Cruz Biotechnology, Inc.).

Techniques: Expressing, Control, Modification, Transfection, shRNA

Journal: Molecular Medicine Reports

Article Title: Effect of Hath1 on the proliferation and apoptosis of cutaneous squamous cell carcinoma in vitro

doi: 10.3892/mmr.2015.4463

Figure Lengend Snippet: Effect of Hath1 on KUMA5 cell growth. Control, blank control group (Dulbecco's modified Eagle's medium/F12); GFP, pGenesil-1 scramble control group; Hath1, pcDNA3.1-Hath1-transfected group; shRNA, pGen-esil-1-Hath1-transfected group. GFP, green fluorescent protein; shRNA, short hairpin RNA.

Article Snippet: The PVDF membranes were then blocked in Tris-buffered saline (TBS; 10 mM Tris-HCl, pH 7.5 and 150 mM NaCl) containing 5% non-fat dry milk at room temperature for 1 h and incubated at 4° C overnight with anti-Hath1 monoclonal antibody (1:400; cat. no. sc-136173; Santa Cruz Biotechnology, Inc.) and anti-β-actin monoclonal antibody (1:1,000; cat. no. sc-47778; Santa Cruz Biotechnology, Inc.).

Techniques: Control, Modification, Transfection, shRNA

Journal: Molecular Medicine Reports

Article Title: Effect of Hath1 on the proliferation and apoptosis of cutaneous squamous cell carcinoma in vitro

doi: 10.3892/mmr.2015.4463

Figure Lengend Snippet: Effect of Hath1 on the apoptosis of KUMA5 cells. Control, blank control group (Dulbecco's modified Eagle's medium/F12); GFP, pGenesil-1 scramble control group; Hath1, pcDNA3.1-Hath1-transfected group; shRNA, pGenesil-1-Hath1-transfected group. GFP, green fluorescent protein; FITC, fluorescein isothiocyanate; shRNA, short hairpin RNA.

Article Snippet: The PVDF membranes were then blocked in Tris-buffered saline (TBS; 10 mM Tris-HCl, pH 7.5 and 150 mM NaCl) containing 5% non-fat dry milk at room temperature for 1 h and incubated at 4° C overnight with anti-Hath1 monoclonal antibody (1:400; cat. no. sc-136173; Santa Cruz Biotechnology, Inc.) and anti-β-actin monoclonal antibody (1:1,000; cat. no. sc-47778; Santa Cruz Biotechnology, Inc.).

Techniques: Control, Modification, Transfection, shRNA

Journal: Cell Death and Differentiation

Article Title: Trim32 suppresses cerebellar development and tumorigenesis by degrading Gli1/sonic hedgehog signaling

doi: 10.1038/s41418-019-0415-5

Figure Lengend Snippet: Trim32 is selectively expressed in inner EGL and displays an uneven cytoplasmic distribution during GNP differentiation. a RT-qPCR analysis of Trim32 in P0, P7, P14, and adult mouse cerebella. Data are expressed as means ± SD (n = 3). An asterisk indicates P < 0.05 and triple asterisks indicate P < 0.001. b Immunoblotting analysis of Trim32, Gli1, and MycN in P0, P7, P14, and adult mouse cerebellums. c Immunofluorescence staining of Trim32 (red) in the EGL of P7 Math1-GFP transgenic mouse cerebellum. Nuclei were counterstained with DAPI (blue). EGL external granule layer, ML molecular layer, PCL Purkinje cells layer, and IGL internal granule layer. The scale bars represent 100 µm in the first panel and 25 µm in the second panel. d Confocal analysis (left) and quantification of immunofluorescence intensities (right) of distribution of Trim32 (green) in the dividing GNPs in different phases of the cell cycle. The dashed line highlights the cells that are in the indicated phase of cell cycle. The cell-cycle phases were identified by PH3 staining (red) for DNA. Nuclei were counterstained with DAPI (blue). The scale bars represent 10 µm. Data are expressed as means ± SD (n = 3). ns indicates P > 0.05 and triple asterisks indicate P < 0.001

Article Snippet: For protein detection, the following antibodies were used: Trim32 (1:1000, GeneTex, #GTX113,937), Gli1 (1:1000, CST, #2643), Ccnd2 (1:1000, CST, #3741), Math1(1:1000, GeneTex, #GTX111,898), NeuN (1:1000, CST, #12,943), Ubiquitin (1:1000, CST, #3936), and actin (1:10,000, CST, #4970).

Techniques: Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining, Transgenic Assay

Journal: Cell Death and Differentiation

Article Title: Trim32 suppresses cerebellar development and tumorigenesis by degrading Gli1/sonic hedgehog signaling

doi: 10.1038/s41418-019-0415-5

Figure Lengend Snippet: Trim32 knockout enhances GNP proliferation in the postnatal developing cerebellum. a Expression of the Math1-GFP protein (GFP, green) in P7 mouse cerebellar sections from the Math1-GFP/Trim32wt mice and Math1-GFP/Trim32KO mice. Nuclei were counterstained with DAPI (blue). Graph in a representing Math1 positive cells normalized to the length of the EGL edge. The scale bar represents 25 µm. Immunofluorescence staining of NeuN (red, b) and Ki67 (red, c) in P7 mouse cerebellar sections from the Math1-GFP/Trim32wt mice and Math1-GFP/Trim32KO mice. Nuclei were counterstained with DAPI (blue). Graph in b and c respectively representing NeuN or Ki67-positive cells normalized to the length of the EGL edge. The scale bar in b represents 50 µm. The scale bar in c represents 25 µm. oEGL outer external granule layer, iEGL inner external granule layer, ML molecular layer, and IGL internal granule layer. d P7 and P18 cerebellar midsagittal sections were stained for DAPI to show the overall morphology of cerebellum in Trim32wt mice and Trim32ko mice. The scale bar represents 500 µm

Article Snippet: For protein detection, the following antibodies were used: Trim32 (1:1000, GeneTex, #GTX113,937), Gli1 (1:1000, CST, #2643), Ccnd2 (1:1000, CST, #3741), Math1(1:1000, GeneTex, #GTX111,898), NeuN (1:1000, CST, #12,943), Ubiquitin (1:1000, CST, #3936), and actin (1:10,000, CST, #4970).

Techniques: Knock-Out, Expressing, Immunofluorescence, Staining

Journal: Cell Death and Differentiation

Article Title: Trim32 suppresses cerebellar development and tumorigenesis by degrading Gli1/sonic hedgehog signaling

doi: 10.1038/s41418-019-0415-5

Figure Lengend Snippet: Trim32 knockout increases the incidence of MB in Ptch1+/− mice. Quantitative PCR mRNA (a; mean ± SD; n = 9) and protein levels (b; n = 3) of Trim32 in mouse medulloblastomas (MB) from Ptch1+/− mice and normal cerebella. c RNA expression level of Trim32 negatively correlated with Gli1 (n = 51) in human SHH MB samples. Data are analyzed from Oncomine database. d Kaplan–Meier analysis of MB incidence in 63 Ptch1+/−/Trim32wt mice (gray line) versus 17 Ptch1+/−/Trim32KO mice (black line). Ptch1+/−/Trim32wt mice and Ptch1+/−/Trim32KO mice, obtained by interbreeding for at least three generations the progeny of Ptch1 heterozygous and Trim32 knock-out mice, were then monitored for the onset of medulloblastoma; (triple asterisks indicate P < 0.001, Logrank test). e Expression of the Math1-GFP protein (GFP, green) in the Math1-GFP mouse medulloblastomas and adjacent cerebellar cortices sections from Ptch1+/−/Trim32wt mice and Ptch1+/−/Trim32KO mice. Nuclei were counterstained with DAPI (blue). The scale bar represents 200 µm. MB medulloblastoma, IGL internal granule layer. RT-qPCR analysis of SHH target genes (f), Trim32 (g), the granule neuronal progenitor marker Maht1 (h), and the differentiated granule cell markers including Tuj1 and NeuN (i) in adult mouse cerebellums and medulloblastomas from Ptch1+/−/Trim32wt mice and Ptch1+/−/Trim32KO mice. Data are expressed as means ± SD (n = 3). An asterisk indicates P < 0.05, double asterisks indicate P < 0.01, and triple asterisks indicate P < 0.001. j Immunoblotting analysis of Trim32, Gli1, Ccnd2, MycN, and NeuN in MB from Ptch1+/−/Trim32wt mice and Ptch1+/−/Trim32KO mice

Article Snippet: For protein detection, the following antibodies were used: Trim32 (1:1000, GeneTex, #GTX113,937), Gli1 (1:1000, CST, #2643), Ccnd2 (1:1000, CST, #3741), Math1(1:1000, GeneTex, #GTX111,898), NeuN (1:1000, CST, #12,943), Ubiquitin (1:1000, CST, #3936), and actin (1:10,000, CST, #4970).

Techniques: Knock-Out, Real-time Polymerase Chain Reaction, RNA Expression, Expressing, Quantitative RT-PCR, Marker, Western Blot

Journal: Cell Death and Differentiation

Article Title: Trim32 suppresses cerebellar development and tumorigenesis by degrading Gli1/sonic hedgehog signaling

doi: 10.1038/s41418-019-0415-5

Figure Lengend Snippet: mRNA expression analysis

Article Snippet: For protein detection, the following antibodies were used: Trim32 (1:1000, GeneTex, #GTX113,937), Gli1 (1:1000, CST, #2643), Ccnd2 (1:1000, CST, #3741), Math1(1:1000, GeneTex, #GTX111,898), NeuN (1:1000, CST, #12,943), Ubiquitin (1:1000, CST, #3936), and actin (1:10,000, CST, #4970).

Techniques: Expressing