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. 2025 Nov 21;13:1716808. doi: 10.3389/fcell.2025.1716808

FIGURE 2.

Panel A displays two microscopic images, with a highlighted area on the left showing sparse cells, and the right showing more dense cell structures. Panel B provides DNA sequencing results comparing WT and CH2-KO, indicating a sequence change. Panel C is a bar graph depicting a significant decrease in CH2 mRNA levels in CH2-KO compared to CON. Panel D shows Western blot results for Fel d1 and β-actin proteins. Panel E is a bar graph indicating a reduction in Fel d1 protein level in CH2-KO versus CON.

Generation of CH2-Knockout Monoclonal Cells. (A) Monoclonal selection of cells after puromycin screening. Scale bar: 100 µm. (B) Sanger sequencing showing a three-base insertion mutation at the target site in the CH2-KO clone. (C) qRT-PCR results for CH2 mRNA expression in control and KO cells. (D) Western blot analysis of Fel d1 protein expression in control and KO cells. (E) Quantification of Fel d1 protein expression levels. Data were obtained from three independent experiments. Data are presented as mean ± standard deviation, and P values were calculated by two-tailed unpaired t-test. Significant differences are indicated by **** (P < 0.0001).