Journal: Stem Cell Research & Therapy

Article Title: Simultaneous generation of transplantable RGC-like and corneal progenitor cells from hiPSCs using a dual-lineage platform

doi: 10.1186/s13287-025-04843-z

Figure Lengend Snippet: Characterization of the cultured hiPSCs derived ITGB4 + corneal cells. A Differentiation of ocular cells from hiPSCs, the representative image showed the various ocular cells, within optic vesicle and the epithelial cells area (black box). Scale bar: 200 μm. B The representative image showed the unsorted epithelial and mesenchymal cells after eliminating the neural cells. C FACS sorted ITGB4 + corneal progenitor cells from hiPSCs derived ocular cells. D Representative images of cultured ITGB4 + corneal progenitor cells. Scale bars: 50 μm. E The single cells clone forming capacity of ITGB4 + corneal progenitor cell. Scale bar: 10 μm. F Immunofluorescent staining of ITGB4 + corneal progenitor cells with ITGB4 (green), Krt14 (red), Krt12 (red), P63 (red), PAX6 (green) and Ki67(green). Scale bars: 100 μm. G The representative image of ITGB4 + corneal progenitor cell spheres. Scale bars: 200 μm. H Immunofluorescent staining of ITGB4 + corneal progenitor cell spheres with ITGB4, Krt14, Krt12 and PAX6. Scale bar: 100 μm. I , J The q-RT PCR results show the genes expression by GAPDH normalized △CT value and fold change

Article Snippet: Krt14 , 1:100 , Novus (NBP2-45032).

Techniques: Cell Culture, Derivative Assay, Staining, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Stem Cell Research & Therapy

Article Title: Simultaneous generation of transplantable RGC-like and corneal progenitor cells from hiPSCs using a dual-lineage platform

doi: 10.1186/s13287-025-04843-z

Figure Lengend Snippet: Generation of hiPSCs derived corneal organoids. A The representative image of hiPSCs derived corneal organoids with transparent property. scale bars: 200 μm. B , C Immunofluorescent staining of corneal organoid sections with ITGB4 (green), Krt14 (red), Krt12 (red) and P63 (red). Scale bars: 100 μm and 50 μm, respectively

Article Snippet: Krt14 , 1:100 , Novus (NBP2-45032).

Techniques: Derivative Assay, Staining

Journal: Cancers

Article Title: Keratin-14 (KRT14) Positive Leader Cells Mediate Mesothelial Clearance and Invasion by Ovarian Cancer Cells

doi: 10.3390/cancers11091228

Figure Lengend Snippet: ( A ) Parallel endpoint Boyden chamber assays. Boyden chamber assays using labelled mesothelial cells overlaid with individual patient-derived ovarian cancer spheroids; we observe no invasion of the mesothelial cells at MALDI imaging collection points; n = 3 wells/sample of one representative experiment. ( B ) Haemotoxylin and Eosin (H&E) staining of the invasive interface. H&E staining identifying the invading interface of ovarian cancer spheroid mesothelial co-cultures and the interface used for MALDI imaging mass spectrometry (IMS). ( C ) MALDI IMS of the invading interface. MALDI IMS identifies: CDCA8, HNRN, keratin-14 (KRT14) and FNDC3B expressed at the invading interface of ovarian/mesothelial co-cultures. ( D ) Representative qRT-PCR of MALDI identified candidates using fresh-frozen confirmed primary high-grade serous ovarian tumours or normal whole ovary ( n = 3/group) where individual data points represent individual patient samples. ( E ) Representative IHC of individual MALDI identified candidates in HGSC primary ovarian samples.

Article Snippet: Cells were transfected with guide strands (1–3), a non-targeting control, or the KRT14 overexpression construct KRT14 OE (Origene, Rockville, United States of America#RC214907) using Lipofectamine ® 2000 Transfection Reagent (Invitrogen, Carlsbad, United States of America #11668019) in DMEM as per the manufacturer’s protocol.

Techniques: Derivative Assay, Imaging, Staining, Mass Spectrometry, Quantitative RT-PCR

Journal: Cancers

Article Title: Keratin-14 (KRT14) Positive Leader Cells Mediate Mesothelial Clearance and Invasion by Ovarian Cancer Cells

doi: 10.3390/cancers11091228

Figure Lengend Snippet: ( A ) Western blot analyses demonstrating the endogenous expression of MALDI identified candidates FNDC3B, HNRN, CDCA8, or KRT14 in ovarian cancer cell lines or the mesothelium LP-9. ( B ) RTCA proliferation assays. Representative results from OVCAR4 real-time proliferation assays conducted in complete culture medium. Data are mean impedance readings from triplicate wells taken every 15 min over a 30 h assay period with lower standard deviation shown, where the non-targeting control is NT and individual CRISPR gene knock-outs are indicated. ( C ) RTCA invasion assays. Representative results from OVCAR4 real-time invasion assays. Data are mean impedance readings from duplicate wells taken every 15 min over a 26 h assay period with lower standard deviation shown. Individual CRISPR gene knock-outs are indicated on the graph. ( D ) Wound-healing assays. Wounded OVCAR4 cell lines (UT = untreated, NT= non-targeting and KRT14 KO = KRT14 CRISPR knockout) were cultured in complete medium. Cells were imaged at regular intervals ranging from 0 to 48 h with the original wound area indicated by dotted lines to aid in the assessment of wound closure. Images presented are representative of the observations of three separate experiments at assay commencement and 48 h, the assay was completed on a minimum of three separate occasions.

Article Snippet: Cells were transfected with guide strands (1–3), a non-targeting control, or the KRT14 overexpression construct KRT14 OE (Origene, Rockville, United States of America#RC214907) using Lipofectamine ® 2000 Transfection Reagent (Invitrogen, Carlsbad, United States of America #11668019) in DMEM as per the manufacturer’s protocol.

Techniques: Western Blot, Expressing, Standard Deviation, Control, CRISPR, Knock-Out, Cell Culture

Journal: Cancers

Article Title: Keratin-14 (KRT14) Positive Leader Cells Mediate Mesothelial Clearance and Invasion by Ovarian Cancer Cells

doi: 10.3390/cancers11091228

Figure Lengend Snippet: ( A ) Representative qRT-PCR of KRT14 expression in LP9 cells, whole normal ovary, benign fibroma, primary ovarian tumour, or ascites-derived cells; and migratory leader cell populations ( n = 3 separate patient isolations where individual data points represent individual patient samples). ( B ) KRT14 expression in cultured cells by immunofluorescence. KRT14 expression was detected as a granular cytoplasmic stain expressed by a small proportion of ovarian cancer cells in the OVCAR4 line cultured as a two-dimensional (2D) monolayer. Scale bar = 100 µm. ( C ) KRT14 expression was detected at the periphery of OVCAR4 ovarian cancer cells cultured as three-dimensional (3D) spheroids with no signal detected in the core of the spheroid. Scale bar = 100 µm. ( D ) Localisation of KRT14 at the periphery of ovarian cancer spheroids by comparison to the intracellular staining observed for N-cadherin. Scale bar = 100 µm. ( E ) Vector control (NT), KRT14 KO , and KRT14 OE cells were suspended in SFM/0.25% methylcellulose in U-bottom 96-well plates and imaged at regular intervals (0–48 h) to observe spheroid aggregation. Images are representative of multiple wells observed at the 13-hour and 48-hour time points. Scale bar = 100 µm. ( F ) Spheroid attachment to the target mesothelium. Vector control (NT) and KRT14 KO spheroids were co-cultured with the target mesothelium, and attachment was measured 6 h post-addition. The results presented are from one representative assay where data points are the attachment counts of three individual wells per cell line.

Article Snippet: Cells were transfected with guide strands (1–3), a non-targeting control, or the KRT14 overexpression construct KRT14 OE (Origene, Rockville, United States of America#RC214907) using Lipofectamine ® 2000 Transfection Reagent (Invitrogen, Carlsbad, United States of America #11668019) in DMEM as per the manufacturer’s protocol.

Techniques: Quantitative RT-PCR, Expressing, Derivative Assay, Cell Culture, Immunofluorescence, Staining, Comparison, Plasmid Preparation, Control

Journal: Cancers

Article Title: Keratin-14 (KRT14) Positive Leader Cells Mediate Mesothelial Clearance and Invasion by Ovarian Cancer Cells

doi: 10.3390/cancers11091228

Figure Lengend Snippet: ( A ) Mesothelial displacement assays. Ovarian cancer spheroids were generated from wild-type (WT), KRT14 KO and KRT14 OE lines and overlaid onto a confluent layer of mesothelial cells and imaged by light microscope. Representative images following overnight and 48-h co-culture are presented where dotted lines indicate spheroid outgrowth and mesothelial displacement. ( B , C ) Embedding and outgrowth assays. OVCAR4 ovarian cancer spheroids were formed and overlaid into ( B ) Collagen I (24 h) or ( C ) Matrigel (48 h) samples, and subsequently fixed and stained for KRT14 expression. ( D ) Wound-healing assays OVCAR4 cells were grown as a monolayer, cells were wounded, fixed, and stained for KRT14 expression at time-points preceding total wound closure. Images from a representative well are shown with scale bars = 100 µm.

Article Snippet: Cells were transfected with guide strands (1–3), a non-targeting control, or the KRT14 overexpression construct KRT14 OE (Origene, Rockville, United States of America#RC214907) using Lipofectamine ® 2000 Transfection Reagent (Invitrogen, Carlsbad, United States of America #11668019) in DMEM as per the manufacturer’s protocol.

Techniques: Generated, Light Microscopy, Co-Culture Assay, Staining, Expressing

Journal: Cancers

Article Title: Keratin-14 (KRT14) Positive Leader Cells Mediate Mesothelial Clearance and Invasion by Ovarian Cancer Cells

doi: 10.3390/cancers11091228

Figure Lengend Snippet: ( A ) Table of the immunohistochemical assessment of KRT14 expression in ovarian cancer tissue microarrays. Staining intensity was scored according to 0 (no stain), 1 (low), 2 (medium), or 3 (high). ( B ) Representative immunohistochemical staining of KRT14 in the various ovarian cancer subtypes (as labelled in figure) from TMA assessments. Scale bar = 200 µm. ( C – F ) Association of KRT14 expression with progression-free survival (PFS) according to ( C ) overall PFS (HR 1.17; 95% CI 1.03–1.33 p < 0.015; B); ( D ) early stage (I/II) diagnosis (HR 1.96; 95% CI 1.08–3.56 p < 0.025); ( E ) following platinum and taxol-based chemotherapy (HR 1.27; 95% CI 1.07–1.51 p < 0.006); and ( F ) following optimal debulk (HR 1.24; 95% CI 1.03–1.5 p < 0.026).

Article Snippet: Cells were transfected with guide strands (1–3), a non-targeting control, or the KRT14 overexpression construct KRT14 OE (Origene, Rockville, United States of America#RC214907) using Lipofectamine ® 2000 Transfection Reagent (Invitrogen, Carlsbad, United States of America #11668019) in DMEM as per the manufacturer’s protocol.

Techniques: Immunohistochemical staining, Expressing, Staining, Biomarker Discovery

Journal: Scientific reports

Article Title: Extracellular C1qbp inhibits myogenesis by suppressing NFATc1.

doi: 10.1038/s41598-024-66549-1

Figure Lengend Snippet: Figure 2. C1qbp expression in skeletal muscles. (A) Total RNAs were isolated from various tissues of male C57BL/6 mice aged 8 weeks. The expression levels of C1qbp, Krt14, and Bax were quantified using qRT-PCR. Muscle indicates gastrocnemius. Data are represented as mean ± SD (n = 3). (B) The expression levels of C1qbp, Krt14, and Bax in various skeletal muscles were examined using qRT-PCR. EDL extensor digitorum longus, TA tibialis anterior. Data are represented as mean ± SD (n = 3). (C) Gastrocnemius muscles of male C57BL/6 mice aged 8 weeks were subjected to immunohistochemical analysis with anti-C1qbp, anti-Krt14, and anti-Bax antibodies. Anti-MyHC and anti-tartrate- resistant acid phosphatase (TRAP) antibodies were used as positive and negative controls, respectively. Representative immunohistochemical images are shown. Scale bar, 100 μm. Arrows indicate the positive signals. (D) The expression level of C1qbp in various skeletal muscle types was analyzed by qRT-PCR. EDL extensor digitorum longus, TA tibialis anterior. Data are represented as mean ± SD (n = 3). **P < 0.01; ***P < 0.001.

Article Snippet: The following antibodies were used in this study: anti-C1qbp (ab24733) and Fast-MyHC (ab91506) antibodies (Abcam, Cambridge, UK); anti-Bax (orb213598), and anti-Krt14 antibodies (orb214169) (Biorbyt, Cambridge, UK); anti-MyCH I (M8421), anti-MyCH II (M1570), anti-Actin (A3854), and anti-tubulin antibodies (T9026) (Sigma, St. Louis, MO, USA); anti-Mef2c (#5030) and anti-histone H3 (#4620) antibodies (Cell Signaling Technology, MA, USA); anti-Lamin A/C (39287) antibody (Thermo Fisher Scientific); anti-TRAP (sc-28204) and anti-NFATc1 antibodies (sc-7294) (Santacruz Biotechnology, Dallas, TX, USA); anti-NFATc3 (GTX133744) (GeneTex, Irvine, CA, USA); anti-p300 (RW105) (Novus Biologicals, USA), and anti-acetyl-histone H3 antibodies (06-599) (Merck, Darmstadt, Germany).

Techniques: Expressing, Muscles, Isolation, Quantitative RT-PCR, Immunohistochemical staining

Journal: iScience

Article Title: Unveiling critical signaling pathways in the murine salivary gland and the role of midkine

doi: 10.1016/j.isci.2026.114852

Figure Lengend Snippet: Branching morphogenesis of embryonic E13.5 SMGs treated with midkine inhibitors is reduced in ex vivo culture (A) Upper panel are light micrographs of wild type C57BL/6J SMGs harvested at E13.5 and cultured for 72 h in the presence of midkine inhibitor (iMDK) or DMSO control. Lower panel shows whole-mount immunofluorescence staining of DMSO control and iMDK treated glands shows expression of Keratin 5 (K5) and neuronal tubulin expressing nerves (Tubb3). Scale bars, 100 μm, ( n = 3). (B) Spooner’s ratio quantification of the number of end buds (expressed as a ratio of the number of end buds at 72 h/number at 1 h) of DMSO control ( n = 10) and iMDK treated glands ( n = 10). (C) Immunofluorescence staining of paraffin embedded DMSO control and iMDK treated glands reveal decreased expression of ΔNp63 and reduced expression of K14 expressing basal stem/progenitor cells. Alterations to the acinar cell differentiation program were also observed in the iMDK treated glands as evident by reduced expression of Mist1, Nkcc1, and Aqp5. Ductal cell differentiation was also altered as depicted by decreased K7 expression in the iMDK treated glands compared to DMSO controls. Scale bars, 20 μm. Lower panel shows quantification of the p63 + K14 + /p63 + basal cells, p63 + Sma + /p63 + MECs, and Nkcc1 + Mist1 + /Nkcc1 + acinar cell population as mean ± SD ( n = 3). (D) Quantitative RT-PCR validation of select genes in the DMSO control and iMDK treated glands. Values were normalized to the housekeeping gene Hprt . Data are represented as means ± SD ( n = 3). ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Primary antibodies used at the indicated dilutions include Midkine (R&D Systems, 1:100), Midkine (ProteinTech, 1:100), K14 (Rizzo et al., 2016, 1:100), Sma (Sigma 1A4, 1:200), Nkcc1 (Santa Cruz Biotechnology, 1:100), ΔNp63 (Cell Signaling Technology, 1:50), ΔNp63 ( ,1:50) Mist1 (Abcam, 1:100), Aqp5 (Alomone Labs, 1:100), Tubb3 (R&D Systems, 1:100), K7 (Abcam, 1:50), K5 (gift from Dr. Julie Segre, 1:100), Troma-III/K19 (Development Studies Hybridoma Bank, 1:50), and Rock1 (Cell Signaling Technology, 1:25).

Techniques: Ex Vivo, Cell Culture, Control, Immunofluorescence, Staining, Expressing, Cell Differentiation, Quantitative RT-PCR, Biomarker Discovery

Journal: Oncology Letters

Article Title: Triple-negative invasive lobular carcinoma presenting with synchronous gastric metastasis: A case report

doi: 10.3892/ol.2025.15242

Figure Lengend Snippet: Immunohistochemistry results of the breast biopsy specimen (magnification, ×100). Staining for (A) ER, (B) PR, (C) HER-2, (D) E-cadherin, (E) Ki-67, (F) β-catenin, (G) CK5/6, (H) CK14 and (I) AR. ER, estrogen receptor; PR, progesterone receptor; CK, cytokeratin; AR, androgen receptor.

Article Snippet: The primary antibodies used included: ER (cat. no. ZA0102), PR (cat. no. ZA-0255), HER-2 (cat. no. ZM-0065), E-cadherin (cat. no. ZA-0565), Ki-67 (cat. no. ZM-0166), CK5/6 (cat. no. ZM-0313), CK14 (cat. no. ZA-0540), AR (cat. no. ZA-0554), GATA-3 (cat. no. ZA-0661), CK7 (cat. no. ZM-0071), CK20 (cat. no. ZA-0574) and Villin (cat. no. ZM-0261), all from Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd. (OriGene Technologies, Inc.) and supplied at a ready to use dilution.

Techniques: Immunohistochemistry, Staining