Fig. 2

NLGP binds to dendritic cells through Dectin-1 receptors. a Widefield fluorescence micrographs of NLGP-FITC bound mouse BMDCs, compared to control (scale bar: 20 µm). b MFIs of experimental replicates (n = 3) of such cell groups as mean ± SD (MFI and CTCF denotes mean fluorescence intensity and corrected total cell fluorescence respectively). c Flow cytometric quantifications of two such representative groups. d Widefield fluorescence micrographs of NLGP-FITC-binding to mBMDCs on neutralizing mentioned cell-surface C-type lectins (scale bar: 20 µm) (MBR denotes mannose binding receptors). e MFIs of experimental replicates (n = 3) of such groups as mean ± SD (TCM denotes tumor conditioning media). f Flow cytometric analyses of these groups. g Agarose gel electrophoresis for reverse transcription-based semi quantitative PCR of IL-10 and IL-12A or IL-12p35 transcripts from control, TCM and TCM + NLGP treated mBMDC groups. h Flow cytometric quantifications of intracellular IL-10 and IL-12p35 from previous groups as offset histograms. i Secretion of IL-10 and IL-12p35 within 48-h culture supernatants of previous groups measured by ELISA, from experimental replicates (n = 6) as mean ± SD. j Agarose gel electrophoresis after reverse transcription-based semi quantitative PCR, measuring IL-10 and IL-12A or IL-12p35 transcripts on neutralizing the mentioned cell-surface C-type lectins on separate groups of TCM + NLGP treated mBMDCs. k Flow cytometric quantifications of intracellular IL-10 and IL-12p35 from receptor-neutralized groups as staggered-offset histograms. l Secretion of IL-10 and IL-12p35, quantified within 48-h culture supernatant of these groups (n = 6) by ELISA as mean ± SD. m Effects of receptor-neutralizations on IL-10 and IL-12A or IL-12p35 production, quantified as transcripts, intracellular proteins, and secreted proteins, compared to control, collectively represented through line diagrams as mean ± SD from (j), (k) and (l). β-actin was taken as loading control in gels. Numerical data of (b), were tested by two-tailed unpaired T test with Welch’s correction (t = 21.52, df = 2.488) and those of (e), (i) and (l) were tested by one-way ANOVA (F = 123.5; 79.20, 96.86; 21.27, 18.09 respectively, df = 20; 17, 17; 41, 41 respectively) followed by Tukey’s multiple comparisons test. Normal distribution of the data were verified before tests (See also Fig. S1)