Journal: Oncogenesis

Article Title: Teriflunomide modulates the PD-1/PD-L1 axis and enhances antitumor immunity in colorectal cancer

doi: 10.1038/s41389-026-00607-3

Figure Lengend Snippet: A CRC cell lines were treated with the indicated concentrations of TER for 72 h. Cell viability assessed using the CCK assay is shown. B Human CRC cell lines were co-cultured with hPD-1 Jurkat-T cells and treated with the indicated concentrations of TER for 72 h. C PD-L1 protein expression in CRC cells co-cultured with hPD-1 Jurkat-T cells and treated with TER for 72 h. GAPDH was used as a loading control. The results are shown as the mean ± SEM. * <0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 compared with the respective control.

Article Snippet: Next, 5 μL of 0.5 mg/mL biotinylated hPD-1 (#71109, BPS Bioscience) was added, and the plates were incubated for 2 h at room temperature.

Techniques: Cell Culture, Expressing, Control

Journal: Oncogenesis

Article Title: Teriflunomide modulates the PD-1/PD-L1 axis and enhances antitumor immunity in colorectal cancer

doi: 10.1038/s41389-026-00607-3

Figure Lengend Snippet: A The viability of hPD-1 Jurkat-T cells and hPD-L1 CHO cells following treatment with the indicated concentrations of TER for 24 h. B Luciferase activity measured using a PD-1/PD-L1 blockade bioassay. hPD-1 Jurkat-T cells (effector cells) were co-cultured with hPD-L1-expressing aAPC/CHO-K1 cells (target cells) in the presence of indicated concentrations of TER. The luminescence signal indicates the level of TCR signaling activation. αPD-L1 was used as a positive control. * <0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 compared with the respective control.

Article Snippet: Next, 5 μL of 0.5 mg/mL biotinylated hPD-1 (#71109, BPS Bioscience) was added, and the plates were incubated for 2 h at room temperature.

Techniques: Luciferase, Activity Assay, Bioassay, Cell Culture, Expressing, Activation Assay, Positive Control, Control

Journal: Oncogenesis

Article Title: Teriflunomide modulates the PD-1/PD-L1 axis and enhances antitumor immunity in colorectal cancer

doi: 10.1038/s41389-026-00607-3

Figure Lengend Snippet: A The viability of hPD-L1 MC38 cells following treatment with the indicated concentrations of TER for 72 h. B CD8 + T cells were isolated from tumors of hPD-1 knock-in mice bearing hPD-L1 MC38 tumors. These tumor-infiltrating CD8 + T cells were co-cultured with hPD-L1 MC38 cells as target cells in the presence of TER for 72 h. Cell viability measured using the CCK assay is depicted. C PD-L1 expression in hPD-L1 MC38 cells, as assessed by western blot analysis using protein lysates from co-culture conditions. GAPDH was used as a loading control. D The levels of immune-related factors, including GrB, IL-2, and IFN-γ, measured in the co-culture supernatant by ELISA. * <0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 compared with the respective control.

Article Snippet: Next, 5 μL of 0.5 mg/mL biotinylated hPD-1 (#71109, BPS Bioscience) was added, and the plates were incubated for 2 h at room temperature.

Techniques: Isolation, Knock-In, Cell Culture, Expressing, Western Blot, Co-Culture Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: Oncogenesis

Article Title: Teriflunomide modulates the PD-1/PD-L1 axis and enhances antitumor immunity in colorectal cancer

doi: 10.1038/s41389-026-00607-3

Figure Lengend Snippet: A Body weight of hPD-1 knock-in mice during the treatment period. The mice were treated with vehicle or TER (10 or 30 mpk) for the indicated time. B Spleen weight of mice at the endpoint of the experiment. C Tumor volume was measured over time in hPD-1 knock-in mice bearing hPD-L1 MC38 tumors treated with vehicle or TER (10 or 30 mpk). Representative images of excised tumors from each group are shown. D Tumor weight at the endpoint of the experiment. E Flow cytometry analysis of CD8 + T-cell populations in tumors from each treatment group. F PD-L1 expression in tumors from each group, as assessed by western blot analysis. GAPDH was used as a loading control. G IHC staining of tumor sections for immune-related markers, including CD8 + T cells and GrB. Representative images from each group are shown, and the quantitation of marker-positive cells per field is presented. * <0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 compared with the respective control.

Article Snippet: Next, 5 μL of 0.5 mg/mL biotinylated hPD-1 (#71109, BPS Bioscience) was added, and the plates were incubated for 2 h at room temperature.

Techniques: Knock-In, Flow Cytometry, Expressing, Western Blot, Control, Immunohistochemistry, Quantitation Assay, Marker

Journal: Oncogenesis

Article Title: Teriflunomide modulates the PD-1/PD-L1 axis and enhances antitumor immunity in colorectal cancer

doi: 10.1038/s41389-026-00607-3

Figure Lengend Snippet: A Body weight of hPD-1 knock-in mice during the treatment period. The mice were treated with vehicle or TER (30 mpk) and received either an isotype control or a CD8 depletion antibody. B Spleen weight of mice at the endpoint of the experiment. C Tumor volume was measured in hPD-1 knock-in mice bearing hPD-L1 MC38 tumors over time following treatment with vehicle or TER (30 mpk) with or without CD8 depletion. Representative images of excised tumors from each group are shown. D Tumor weight at the endpoint of the experiment. E Flow cytometry analysis confirming CD8 + T-cell depletion in tumors from each treatment group. The proportion of CD8 + cells among total live cells was quantified. F IHC staining of tumor sections for CD8 + T cells and GrB. Representative images from each treatment group are shown, and the quantitation of marker-positive cells per field is presented. * <0.05, ** p < 0.01, and **** p < 0.0001 compared with the respective control.

Article Snippet: Next, 5 μL of 0.5 mg/mL biotinylated hPD-1 (#71109, BPS Bioscience) was added, and the plates were incubated for 2 h at room temperature.

Techniques: Knock-In, Control, Flow Cytometry, Immunohistochemistry, Quantitation Assay, Marker

Journal: Molecular Cancer

Article Title: Activation of the hedgehog pathway in advanced prostate cancer

doi: 10.1186/1476-4598-3-29

Figure Lengend Snippet: Detection of HIP in human cancer specimens. By Western blotting, HIP antibodies (R&D systems Cat# AF1568) recognized one band between 75 and100 KD ( A ). Expression of endogenous HIP was detected in two GI cancer tissues, which were known to contain activated hedgehog signaling (data not shown here), but not in the matched normal tissue ( B ). Immunohistostaining of HIP I prostate cancer showed a similar pattern to PSA ( C , 200×)

Article Snippet: A standard avidin-biotin immunostaining technique was performed using a kit from Vector laboratories using specific antibodies to Su(Fu) (Santa Cruz Biotechnology Cat# 10933), PTCH1 (Santa Cruz Biotechnology Cat# 6149), HIP (R&D systems Cat# AF1568) and Shh (Santa Cruz Biotechnology Cat# 9024) and PSA (Vector laboratories).

Techniques: Western Blot, Expressing

Journal: bioRxiv

Article Title: Human alveolar Type 2 epithelium transdifferentiates into metaplastic KRT5+ basal cells during alveolar repair

doi: 10.1101/2020.06.06.136713

Figure Lengend Snippet: (A) IPA upstream regulator analysis of hAEC2s co-cultured with AHLM. (B) Bulk RNA-seq analysis of MRC5 and AHLM with differential gene expression analysis. (C) Treatment of hAEC2s+AHLM organoids with BMP4 increases SFTPC and decreases KRT5 mRNA expression at D14 of culture. (D) BMP treatment increases SFTPC+ cells and decreases KRT5+ cells in D14 organoids. (E) hAEC2s from IPF lungs demonstrates decreased SFTPC and HHIP expression when compared to hAEC2s from normal donors. (F) Treatment of hAEC2s+AHLM organoids with HHIP increases SFTPC and decreases KRT5 mRNA expression at D14 of culture. (G) HHIP increases SFTPC+ cells and decreases KRT5+ cells in D14 organoids. (H) HHIP-treated AHLM isolated from D14 organoid culture have decreased GLI1 and increased BMP3 and BMP4 mRNA levels. (I) HHIP-treated hAEC2s+AHLM organoids at D14 show significantly higher fraction of pSMAD1/5/8+ cells. See also

Article Snippet: Where applicable, recombinant BMP4 (Cat#314-BP-010; R&D Systems; used 50 ng/mL) and HHIP (Cat#9280-HP-050; R&D Systems; used 5 μg/mL) were added to the media after 48 hours and replenished in every media change.

Techniques: Cell Culture, RNA Sequencing, Gene Expression, Expressing, Isolation

Journal: Scientific Reports

Article Title: Podocalyxin promotes the formation of compact and chemoresistant cancer spheroids in high grade serous carcinoma

doi: 10.1038/s41598-024-57053-7

Figure Lengend Snippet: Analysis of spheroids formed with representative ovarian cancer cell lines. The four cell lines shown in Fig. B (Kuramochi, HEY, SKOV3 and COV362) were analysed. Bright field images and immunofluorescent staining for PODXL (green), nuclei (blue) and F-actin (red) are shown at 20× magnification. Scale bar: 100 µm.

Article Snippet: Spheroids were fixed with 4% paraformaldehyde in PBS for 30 min, permeabilised with 0.1% Triton X-100 in PBS for 10 min, and blocked with 15% normal donkey serum in 1% BSA/PBS for 2 h. Spheroids were incubated overnight at 4 °C with goat anti-human PODXL antibodies (2.5 μg/ml diluted in 1% BSA/PBS, R&D Systems, #AF1568) or goat IgG (2.5 μg/ml diluted in 1% BSA/PBS, R&D Systems, #AB-108-C), then 2 h with Alexa Fluor 488 donkey anti-goat antibodies (1 μg/ml diluted in 1% BSA/PBS, Invitrogen, #A11055).

Techniques: Staining

Journal: Scientific Reports

Article Title: Podocalyxin promotes the formation of compact and chemoresistant cancer spheroids in high grade serous carcinoma

doi: 10.1038/s41598-024-57053-7

Figure Lengend Snippet: Validation of PODXL-KO in Kuramochi cell line. PODXL was knocked out in Kuramochi and controls were transfected with an empty vector. ( A ) Real-time RT-PCR analysis of PODXL mRNA. Data normalized to 18S and expressed as fold changes relative to control. ( B ) Western blot analysis of PODXL protein, β-actin used as a loading control. Left panel: a representative image. Right panel: densitometric analysis of the top band of PODXL, data normalized to β-actin and expressed as fold changes relative to control (full western blot is presented in Supplementary Fig. ). ( C ) Representative images of spheroids formed with control and PODXL-KO cells. Top: bright field. Images at 4× magnification. Bottom: immunofluorescent staining of PODXL (green), nuclei (blue), and F-actin (red). Images at 20× magnification. Scale bar: 100 µm. Mean ± SD, n = 3. **P < 0.005, ****P < 0.0001.

Article Snippet: Spheroids were fixed with 4% paraformaldehyde in PBS for 30 min, permeabilised with 0.1% Triton X-100 in PBS for 10 min, and blocked with 15% normal donkey serum in 1% BSA/PBS for 2 h. Spheroids were incubated overnight at 4 °C with goat anti-human PODXL antibodies (2.5 μg/ml diluted in 1% BSA/PBS, R&D Systems, #AF1568) or goat IgG (2.5 μg/ml diluted in 1% BSA/PBS, R&D Systems, #AB-108-C), then 2 h with Alexa Fluor 488 donkey anti-goat antibodies (1 μg/ml diluted in 1% BSA/PBS, Invitrogen, #A11055).

Techniques: Biomarker Discovery, Transfection, Plasmid Preparation, Quantitative RT-PCR, Control, Western Blot, Staining

Journal: Scientific Reports

Article Title: Podocalyxin promotes the formation of compact and chemoresistant cancer spheroids in high grade serous carcinoma

doi: 10.1038/s41598-024-57053-7

Figure Lengend Snippet: Analysis of growth and spheroid compactness of control and PODXL-KO Kuramochi cells. Control and PODXL-KO spheroids were monitored for 24, 48 and 72 h. ( A ) Representative brightfield images at 4× magnification. Scale bar: 50 µm. ( B ) Cell viability. ( C ) Total number of live cells. ( D ) Spheroid compactness. ( E ) Spheroid hardiness. Data expressed as percentage of intact spheroids over total number of spheroids. Mean ± SD, n = 3. *P < 0.05, **P < 0.005.

Article Snippet: Spheroids were fixed with 4% paraformaldehyde in PBS for 30 min, permeabilised with 0.1% Triton X-100 in PBS for 10 min, and blocked with 15% normal donkey serum in 1% BSA/PBS for 2 h. Spheroids were incubated overnight at 4 °C with goat anti-human PODXL antibodies (2.5 μg/ml diluted in 1% BSA/PBS, R&D Systems, #AF1568) or goat IgG (2.5 μg/ml diluted in 1% BSA/PBS, R&D Systems, #AB-108-C), then 2 h with Alexa Fluor 488 donkey anti-goat antibodies (1 μg/ml diluted in 1% BSA/PBS, Invitrogen, #A11055).

Techniques: Control

Journal: Scientific Reports

Article Title: Podocalyxin promotes the formation of compact and chemoresistant cancer spheroids in high grade serous carcinoma

doi: 10.1038/s41598-024-57053-7

Figure Lengend Snippet: Comparison of control and PODXL-KO Kuramochi spheroids following treatment with carboplatin. ( A – C ) Control and PODXL-KO spheroids were treated with carboplatin for 24 h, then monitored for up to 4 days. ( A ) Representative brightfield images of carboplatin treated spheroids in recovery. Scale bar: 100 µm. ( B ) Cell viability. ( C ) Total number of live cells within each spheroid. ( D ) Immunofluorescent staining of Ki-67 in spheroids at day 0 (immediately after carboplatin treatment). Left panel: representative images at 40× magnification. Scale bar: 50 µm. Right panel: percentage of Ki-67-positive cells Mean ± SD, n = 3. *P < 0.05, **P < 0.005.

Article Snippet: Spheroids were fixed with 4% paraformaldehyde in PBS for 30 min, permeabilised with 0.1% Triton X-100 in PBS for 10 min, and blocked with 15% normal donkey serum in 1% BSA/PBS for 2 h. Spheroids were incubated overnight at 4 °C with goat anti-human PODXL antibodies (2.5 μg/ml diluted in 1% BSA/PBS, R&D Systems, #AF1568) or goat IgG (2.5 μg/ml diluted in 1% BSA/PBS, R&D Systems, #AB-108-C), then 2 h with Alexa Fluor 488 donkey anti-goat antibodies (1 μg/ml diluted in 1% BSA/PBS, Invitrogen, #A11055).

Techniques: Comparison, Control, Staining

Journal: Scientific Reports

Article Title: Podocalyxin promotes the formation of compact and chemoresistant cancer spheroids in high grade serous carcinoma

doi: 10.1038/s41598-024-57053-7

Figure Lengend Snippet: Analysis of cancer spheroids formed with ascites-derived cells from HGSC patients. ( A ) Bright field images of spheroids formed from 6 independent samples examined and their immunofluorescent staining of PODXL (green), nuclei (blue), and F-actin (red). Images at 4 and 20× magnification respectively. ( B – F ) Further analysis of cells expressing the highest (#1) and lowest (#6) levels of PODXL. ( B ) Representative brightfield images of spheroids formed over time. Images at 4× magnification. ( C ) Cell viability. ( D ) Total number of live cells. ( E ) Spheroid compactness. ( F ) Spheroid hardiness. Data expressed as percentage of intact spheroids over total number of spheroids. Scale bar: 100 µm. Mean ± SD, n = 3. *P < 0.05, **P < 0.005, ***P < 0.001.

Article Snippet: Spheroids were fixed with 4% paraformaldehyde in PBS for 30 min, permeabilised with 0.1% Triton X-100 in PBS for 10 min, and blocked with 15% normal donkey serum in 1% BSA/PBS for 2 h. Spheroids were incubated overnight at 4 °C with goat anti-human PODXL antibodies (2.5 μg/ml diluted in 1% BSA/PBS, R&D Systems, #AF1568) or goat IgG (2.5 μg/ml diluted in 1% BSA/PBS, R&D Systems, #AB-108-C), then 2 h with Alexa Fluor 488 donkey anti-goat antibodies (1 μg/ml diluted in 1% BSA/PBS, Invitrogen, #A11055).

Techniques: Derivative Assay, Staining, Expressing

Journal: Scientific Reports

Article Title: Podocalyxin promotes the formation of compact and chemoresistant cancer spheroids in high grade serous carcinoma

doi: 10.1038/s41598-024-57053-7

Figure Lengend Snippet: Analysis of cancer spheroids of primary HGSC cells following treatment with carboplatin. Cells expressing the highest (#1) and lowest (#6) levels of PODXL shown in Fig. A were analysed. ( A – C ) Spheroids were treated with carboplatin for 24 h then monitored for up to 4 days. ( A ) Representative brightfield images of carboplatin treated spheroids in recovery. Images at 4× magnification. ( B ) Cell viability. ( C ) Total Number of live cells within each spheroid. ( D ) Immunofluorescent staining of Ki-67 in spheroids at day 0 (immediately after carboplatin treatment). Representative images at 20× magnification. Scale bar: 100 µm. Mean ± SD, n = 3. *P < 0.05, **P < 0.005.

Article Snippet: Spheroids were fixed with 4% paraformaldehyde in PBS for 30 min, permeabilised with 0.1% Triton X-100 in PBS for 10 min, and blocked with 15% normal donkey serum in 1% BSA/PBS for 2 h. Spheroids were incubated overnight at 4 °C with goat anti-human PODXL antibodies (2.5 μg/ml diluted in 1% BSA/PBS, R&D Systems, #AF1568) or goat IgG (2.5 μg/ml diluted in 1% BSA/PBS, R&D Systems, #AB-108-C), then 2 h with Alexa Fluor 488 donkey anti-goat antibodies (1 μg/ml diluted in 1% BSA/PBS, Invitrogen, #A11055).

Techniques: Expressing, Staining

Journal: Oncogenesis

Article Title: Teriflunomide modulates the PD-1/PD-L1 axis and enhances antitumor immunity in colorectal cancer

doi: 10.1038/s41389-026-00607-3

Figure Lengend Snippet: A The viability of hPD-1 Jurkat-T cells and hPD-L1 CHO cells following treatment with the indicated concentrations of TER for 24 h. B Luciferase activity measured using a PD-1/PD-L1 blockade bioassay. hPD-1 Jurkat-T cells (effector cells) were co-cultured with hPD-L1-expressing aAPC/CHO-K1 cells (target cells) in the presence of indicated concentrations of TER. The luminescence signal indicates the level of TCR signaling activation. αPD-L1 was used as a positive control. * <0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 compared with the respective control.

Article Snippet: Recombinant hPD-L1 (#71104, BPS Bioscience) was coated onto 96-well plates (Corning Inc., New York, NY, USA) at a concentration of 1 mg/mL in PBS and incubated overnight.

Techniques: Luciferase, Activity Assay, Bioassay, Cell Culture, Expressing, Activation Assay, Positive Control, Control

Journal: Oncogenesis

Article Title: Teriflunomide modulates the PD-1/PD-L1 axis and enhances antitumor immunity in colorectal cancer

doi: 10.1038/s41389-026-00607-3

Figure Lengend Snippet: A The viability of hPD-L1 MC38 cells following treatment with the indicated concentrations of TER for 72 h. B CD8 + T cells were isolated from tumors of hPD-1 knock-in mice bearing hPD-L1 MC38 tumors. These tumor-infiltrating CD8 + T cells were co-cultured with hPD-L1 MC38 cells as target cells in the presence of TER for 72 h. Cell viability measured using the CCK assay is depicted. C PD-L1 expression in hPD-L1 MC38 cells, as assessed by western blot analysis using protein lysates from co-culture conditions. GAPDH was used as a loading control. D The levels of immune-related factors, including GrB, IL-2, and IFN-γ, measured in the co-culture supernatant by ELISA. * <0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 compared with the respective control.

Article Snippet: Recombinant hPD-L1 (#71104, BPS Bioscience) was coated onto 96-well plates (Corning Inc., New York, NY, USA) at a concentration of 1 mg/mL in PBS and incubated overnight.

Techniques: Isolation, Knock-In, Cell Culture, Expressing, Western Blot, Co-Culture Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: Oncogenesis

Article Title: Teriflunomide modulates the PD-1/PD-L1 axis and enhances antitumor immunity in colorectal cancer

doi: 10.1038/s41389-026-00607-3

Figure Lengend Snippet: A Body weight of hPD-1 knock-in mice during the treatment period. The mice were treated with vehicle or TER (10 or 30 mpk) for the indicated time. B Spleen weight of mice at the endpoint of the experiment. C Tumor volume was measured over time in hPD-1 knock-in mice bearing hPD-L1 MC38 tumors treated with vehicle or TER (10 or 30 mpk). Representative images of excised tumors from each group are shown. D Tumor weight at the endpoint of the experiment. E Flow cytometry analysis of CD8 + T-cell populations in tumors from each treatment group. F PD-L1 expression in tumors from each group, as assessed by western blot analysis. GAPDH was used as a loading control. G IHC staining of tumor sections for immune-related markers, including CD8 + T cells and GrB. Representative images from each group are shown, and the quantitation of marker-positive cells per field is presented. * <0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 compared with the respective control.

Article Snippet: Recombinant hPD-L1 (#71104, BPS Bioscience) was coated onto 96-well plates (Corning Inc., New York, NY, USA) at a concentration of 1 mg/mL in PBS and incubated overnight.

Techniques: Knock-In, Flow Cytometry, Expressing, Western Blot, Control, Immunohistochemistry, Quantitation Assay, Marker

Journal: Oncogenesis

Article Title: Teriflunomide modulates the PD-1/PD-L1 axis and enhances antitumor immunity in colorectal cancer

doi: 10.1038/s41389-026-00607-3

Figure Lengend Snippet: A Body weight of hPD-1 knock-in mice during the treatment period. The mice were treated with vehicle or TER (30 mpk) and received either an isotype control or a CD8 depletion antibody. B Spleen weight of mice at the endpoint of the experiment. C Tumor volume was measured in hPD-1 knock-in mice bearing hPD-L1 MC38 tumors over time following treatment with vehicle or TER (30 mpk) with or without CD8 depletion. Representative images of excised tumors from each group are shown. D Tumor weight at the endpoint of the experiment. E Flow cytometry analysis confirming CD8 + T-cell depletion in tumors from each treatment group. The proportion of CD8 + cells among total live cells was quantified. F IHC staining of tumor sections for CD8 + T cells and GrB. Representative images from each treatment group are shown, and the quantitation of marker-positive cells per field is presented. * <0.05, ** p < 0.01, and **** p < 0.0001 compared with the respective control.

Article Snippet: Recombinant hPD-L1 (#71104, BPS Bioscience) was coated onto 96-well plates (Corning Inc., New York, NY, USA) at a concentration of 1 mg/mL in PBS and incubated overnight.

Techniques: Knock-In, Control, Flow Cytometry, Immunohistochemistry, Quantitation Assay, Marker