(A–C) Statistical comparison for the substrate stiffness discrimination capability of B cells from the following three groups: (A) DT40-WT, DT40-Lyn-KO, and DT40-Lyn-Rescue cells; (B) DT40-WT, DT40-Syk-KO, and DT40-Syk-Rescue cells; (C) DT40-WT, DT40-PLCγ2-KO, and DT40-PLCγ2-Rescue cells. The calculation of ratio of BCR MFI is defined as the BCR MFI of each cell on stiff substrates to the averaged value of the BCR MFI of all the cells on soft substrates. (D–F) B1-8 primary B cell pre-treatment with DMSO as a control (NC) versus Lyn inhibitor PP2 (D), Syk inhibitor Piceatannol (E), and PLCγ2 inhibitor U73122 (E). (G–I) CH27 B cell pre-treatment with DMSO as a control (NC) versus Lyn inhibitor PP2 (G), Syk inhibitor Piceatannol (H) and PLCγ2 inhibitor U73122 (I). In (A) to (I), bar represents mean ± SEM from at least 20 cells in one representative of three independent experiments. Two-tailed t tests were performed for statistical comparisons.