Fig. 1: Blimp1 is induced in the tumoral region of the lung from patients with lung adenocarcinoma (LUAD).

a Representative Blimp1 immunofluorescence (IF) staining on histologic sections in the lung from a control subject (CN) and from a patient with lung adenocarcinoma (ADC). TU tumoral region. Higher expression of Blimp1 (in green) was noted in the TU region of patients with lung adenocarcinoma (ADC) as compared to a control (CN) subject. Blimp1+ cells are green in the image from the lung of a CN subject (left panel) and from the tumoral region of the lung of a NSCLC patient with tumor grade G2 having a tumor diameter of 4.8 cm, stage IIA at surgery (right panel). The nuclei are counterstained in blue with DAPI staining on the mounting medium. b Double IF for Blimp1 and CD3 showing co-localization in the tumoral region of the lung of patients with NSCLC. Blimp-1 positive cells (green) were co-localized in the lung tumor microenvironment with CD3+ positive T cells (red stained). Co-localization of CD3+ (red) with Blimp1 (green nuclei) is shown. Nuclei were counterstained with DAPI. Pictures’ magnifications are indicated below the microscopic picture. (MP Molecular Pneumology) MP-3 = subject 3 Grading G3 and stadium IIA having a tumor of 5 cm in diameter at surgery (Table S1). In the right picture, the same staining in a patient with NSCLC, Grading G3 and Stage IVA, with a tumor diameter = 1.3cm at surgery. c A computer tomography of a human right thorax with lung non-small cell lung cancer. Schematic drawing of the regions where tissue samples were collected and dissected for further analysis: tumoral area (TU) (red circle), peritumoral area (PT) (2 cm around the tumor) (yellow circle), and control region (CTR) (>5 cm away from the tumor). d Immediately after lung surgery, lung samples from control subject CN = lung and from NSCLC patients (histologically classified as ADC) were dissected from the tumor area (TU), the peri-tumoral area (PT) surrounding the tumor, and from the control area (CTR). Western blot analysis of total proteins extracted from NSCLC lung tissue samples including CN (patients without tumor), CTR (control area), PT area and TU (tumoral area) is shown. Luminescence-mediated analysis using a polyclonal antibody against Blimp1 was performed, and the Blimp1 protein was detected at around 98 kDa, and GAPDH at 36 kDa. e Western blot analyses for Blimp-1 in lung tissues were normalized for total sample proteins and are shown as mean values ± SEM. Expression Blimp1 protein analysis of the 3 western blots from the top left to right (e–h), respectively. Abbreviations: Carcinoma histologic grading G1 (Low grade): well-differentiated—cells look similar to normal and tend to grow slowly. Specifically for Adenocarcinoma G1: ≤20% high-grade histologic patterns (lepidic predominant); G2 (Intermediate grade): Moderately differentiated—cells have noticeable abnormalities and a moderate growth rate. Adenocarcinoma : G2: 5–50% intermediate patterns (acinar/papillary); G3 (High grade): Poorly differentiated—cells look very abnormal and often grow quickly; adenocarcinoma:G3: >20% high-grade patterns (solid/micropapillary predominant). MT metastasis. i Analysis of Blimp1/GAPDH mRNA extracted from the lung of subjects with NSCLC and from the CTR, PT, and TU regions of patients with NSCLC. Detailed single results are reported in Supplementary data. Like Blimp1(i), also T-bet (j) and Perforin/GAPDH (k), CD8/GAPDH (l), and CD4/GAPDH (m) mRNA expression were found to be downregulated in the tumoral region of the lung as compared to the peri-tumoral region of the lung in NSCLC patients. n, o However, the ratio Blimp-1/CD8 and Blimp-1/CD4 mRNA was not downregulated, and the ratio Blimp-1/CD8 was even found to be significantly upregulated in the tumoural region (CN: n = 3; groups of ADC patients: n = 13–16). Data are shown as mean values ± SEM using Student's two-tailed t test. *p < 0.05; **p < 0.01; ***p < 0.001.